Marker In Acute Lymphoblastic Leukemia Research Articles

Aberrant DNA methylation of CDKN2B gene in the Type 1 diabetes mellitus and analysis of epigenetic markers

IntroductionAberrant DNA methylation is one of the important molecular markers in acute lymphoblastic leukemia (ALL). ObjectivesTo determine Aberrant DNA methylation profiles of the CDKN2A and CDKN2B genes in ALL samples from T1DM cases. ResultsAberrant hypermethylation of CDKN2B gene and cytogenetic abnormalities were analyzed in >60% of subjected cases. Total cholesterol, triglycerides and low-density lipoprotein cholesterol were not significantly varied between experimental and control (p >0.05). Expression analysis of human peripheral blood mononuclear cells revealed decreased mRNA levels of CDKN2B. The decreased level of p21 and CDK4 expression in the T1DM cases than control. The number of monocytes, lymphocytes and neutrophils significantly varied in T1DM samples and control (p < 0.05). ConclusionsAberrant methylation is one of the important mechanisms involved in the expression of CDKN2B genes and is an important prognostic marker for the early detection of ALL risks.

Prognostic Value of Serum Paraprotein Response Kinetics in Patients With Newly Diagnosed Multiple Myeloma.

Response kinetics is a well-established prognostic marker in acute lymphoblastic leukemia. The situation is not clear in multiple myeloma (MM) despite having a biomarker for response monitoring (monoclonal component [MC]). We developed a mathematical model to assess the prognostic value of serum MC response kinetics during 6 induction cycles, in 373 NDMM transplanted patients treated in the GEM2012Menos65 clinical trial. The model calculated a "resistance" parameter that reflects the stagnation in the response after an initial descent. Two patient subgroups were defined based on low and high resistance, that respectively captured sensitive and refractory kinetics, with progression-free survival (PFS) at 5 years of 72% and 59% (HR 0.64, 95% CI 0.44-0.93; P=.02). Resistance significantly correlated with depth of response measured after consolidation (80.9% CR and 68.4% minimal residual disease negativity in patients with sensitive vs. 31% and 20% in those with refractory kinetics). Furthermore, it modulated the impact of reaching CR after consolidation; thus, within CR patients those with refractory kinetics had significantly shorter PFS than those with sensitive kinetics (median 54 months vs. NR; P=.02). Minimal residual disease negativity abrogated this effect. Our study also questions the benefit of rapid responders compared to late responders (5-year PFS 59.7% vs. 76.5%, respectively [P < .002]). Of note, 85% of patients considered as late responders were classified as having sensitive kinetics. This semi-mechanistic modeling of M-component kinetics could be of great value to identify patients at risk of early treatment failure, who may benefit from early rescue intervention strategies.

Prognostic significance of aberrant expression of CD markers in acute lymphoblastic leukemia

Acute lymphoblastic leukemia (ALL) forms a heterogeneous group of disorders with a characteristic set of CD markers. Nowadays, these markers are used for the classification of hematopoietic malignancies and are a basis for diagnosis of leukemia. Unusual expression of CD markers as different immunophenotype of lymphoblasts is known as the aberrant expression of markers. The frequency of this phenomenon has been reported to be nearly 30% in different studies. Immunophenotype of precursor B‑cell leukemia involves the likely expression of myeloid CD markers (CD13 and CD33). In precursor T‑cell leukemia, myeloid markers (CD13 and CD33) are frequent but CD117 is rare. On the other hand, the presence of multiple translocations is also associated with a higher or lower level of aberrant markers that could be effective in changing the prognosis of patients. Further studies are needed to confirm the relationship between aberrant expression with prognosis and response to treatment of ALL.

Fat1 cadherin provides a novel minimal residual disease marker in acute lymphoblastic leukemia

Measurement of minimal residual disease (MRD) maintains an important role in the clinical management of acute lymphoblastic leukemia (ALL). Recently, we identified Fat1 cadherin as a unique and independent prognostic factor for relapse-free and overall survival in pediatric pre-B-ALL. Here, we analyzed Fat1 mRNA for its potential as a novel marker of MRD in cases of pre-B- and T-ALL. Analyses of microarray data from 125 matched diagnosis/relapse samples across three independent datasets indicate that Fat1 mRNA is detectable in an average of 31.3% of diagnosed pre-B-ALL, of which 67.5% of cases remained positive at relapse. Furthermore, some 20% of cases with undetectable levels of Fat1 mRNA at diagnosis became positive upon relapse. T-ALL cases were 83.3% positive for Fat1 expression at diagnosis with 77.7% remaining positive at relapse. Towards proof of concept, we developed a quantitative polymerase chain reaction assay and demonstrate detection of Fat1 mRNA in leukemic cells mixed with normal peripheral blood cells at a sensitivity of 1 in 10 000 to 100 000 cells. Fat1 may therefore provide a new marker of MRD for patients with ALL lacking known genomic aberrations or within a multiplex approach to MRD detection.

ANALYSIS OF THE NUMBER OF CELLS WITH CD34+CD38- AND CD34+CD38-CD19+ PHENOTYPES AS POTENTIAL LEUKEMIC STEM CELLS IN ACUTE LYMPHOBLASTIC LEUKEMIA IN CHILDREN

The number of the supposed leukemic stem cells in marrow samples of 54 patients with primary B-linear acute lymphoblastic leukemia was detected by the method of multiparametric flow cytofluorimetry in leukemia diagnosis (zero day). The level of minimal residual disease was estimated on zero and on the fifteenth days of induction therapy. In the course of the research it was found out that leukemic B-cell precursors with СD34+СD38-CD19+ phenotype prevailed among the cells with СD34+СD38-phenotype. The high percentage of both СD34+СD38-, СD34+СD38-, and СD34+СD38-CD19+ among the general population leukemic cells was associated with a worse response to the therapy. Therefore, the initial number of such cells can be considered as a prognostic marker in acute lymphoblastic leukemia in children.

Altered expression of Fat1 cadherin, a novel tumor marker for acute lymphoblastic leukemia

Altered expression of Fat1 cadherin, a novel tumor marker for acute lymphoblastic leukemia

Expression of functional markers in acute lymphoblastic leukemia

We analyzed surface antigens, multidrug resistance (MDR) parameters (PGP, MRP, LRP), tissue infiltration parameters (CD18, CD44, VCAM, MMP2), receptors for colony stimulating factors (G-CSFr, GM-CSFr) and cell cycle parameters (Ki-67, topoisomerase IIα) in 86 patients with acute lymphoblastic leukemia (ALL). LRP, PGP and CD18 were associated with poor clinical outcome, and LRP expression was related with CD18, CD44 and G-CSFr. Of the cell cycle parameters, Ki-67 (+) fraction was increased in ALL with hepato-splenomegaly and extramedullary involvement. In conclusion, analysis of LRP, PGP, CD18 and Ki-67 could be helpful to predict the clinical behavior of ALL.

Cytogenetic Abnormalities and Molecular Markers of Acute Lymphoblastic Leukemia

The chromosomal abnormalities that occur in acute lymphoblastic leukemia have greatly elucidated the biologic causes (leukemogenesis) of this disease. ALL is extremely heterogeneous; different cell types at differing stages of differentiation may become leukemic. Although not yet completely understood, the chromosomal abnormalities occurring in this disease are not random. Rather, the abnormalities are highly specific, with specific abnormalities occurring in specific cell phenotypes. Early chromosomal investigations were concerned with identification of heterogeneous cytogenetic "groups," with independent prognostic indications, among patients with ALL. These studies progressed to the identification of specific chromosomal translocations, many of which can be correlated with specific cell phenotypes. Molecular studies of these cytogenetic aberrations have defined specific chromosomal breakpoints, and specific genes, which have been implicated in the oncogenic process. As a result of these translocations, the involved genes either produce abnormal products or are deregulated. The products of these genes vary, but all are concerned with cell growth and differentiation. Much remains to be determined regarding cytogenetic abnormalities in acute lymphoblastic leukemia and their clinical and biologic significance. The molecular bases of all of the recurring abnormalities in ALL have not been determined. The biologic causes for the chromosomal abnormalities are not known, nor is it known why specific chromosomal aberrations correlate with prognosis.

Possibilities and Limitations of Immunological Marker Analyses for the Detection of Minimal Residual Disease in Childhood Acute Lymphoblastic Leukemia

Specific application of immunological markers in acute lymphoblastic leukemia (ALL) for detection of inadequate blast cell reduction after induction therapy or early recognition of a relapse requires a precise characterization of the immunophenotype at the time of diagnosis and an understanding of the biology of the disease. Therefore, the ALL-BFM 83 study is first used to demonstrate the incidence, antigen expression and dynamics of relapse occurrence of immunological subtypes in childhood ALL. This is then followed by a discussion of the different immunological features hitherto applied for identification of residual leukemia cells in ALL (terminal deoxynucleotidyl transferase--TdT; common acute lymphoblastic leukemia-associated antigen--CALLA, CD10; various T-cell differentiation antigens; kappa/lambda labelling); these are, however, not leukemia-specific and are expressed to varying degrees by normal lymphoid progenitor cells. The sensitivity of these analyses is therefore largely determined by the markers applied and the type of investigational material. Finally, suitable markers are presented for detecting residual leukemia cells in the different immunological subtypes of ALL. Their clinical relevance still remains to be evaluated in prospective therapy studies.

Immunogold staining in bone marrow aspirate for the study of surface markers in acute lymphoblastic leukemia

The diagnostic value of phenotyping acute lymphoblastic leukemia (ALL) of childhood using the immunogold staining method (IGS) was assessed in 34 consecutive new ALL patients seen at Children's Hospital of Pittsburgh during a 14-month period. IGS proved to have considerable value in the diagnosis and classification of ALL. It allowed a sharp and clear identification of surface antigens, documented surface antigen heterogeneity in normal and abnormal cells, and provided a permanent record. The combination with cytochemistries allowed further cytoplasmic identification and correlation. This method does not require sophisticated equipment and can be performed in any laboratory, thus enabling results from different institutions to be more readily compared.

Lineage-related polypeptide markers in acute lymphoblastic leukemia detected by two-dimensional gel electrophoresis.

We have utilized two-dimensional polyacrylamide gel electrophoresis, coupled with ultrasensitive silver staining, to identify lineage-related polypeptide markers in lymphoblasts from children with acute lymphoblastic leukemia. Twelve polypeptides were detected that could distinguish between the major subgroups of acute lymphoblastic leukemia. These included a new marker for common acute lymphoblastic leukemia and markers for cells of B and T lineages. Analysis of the two-dimensional patterns also allowed the tentative identification of T-cell lineage in two cases with an otherwise undifferentiated non-T-cell non-B-cell phenotype. Two-dimensional electrophoresis thus provides a powerful tool for the delineation of the cell of origin in leukemia.

Characterization of a new chromosomal marker for acute lymphoblastic leukemia from a long-term cell line

A bone marrow aspirate from a child with acute lymphoblastic leukemia (ALL) at first relapse was used to establish cell line # 697. The cultured line and marrow aspirates taken at initial diagnosis and first relapse were examined and compared. Similarities in all patterns evaluated confirmed the leukemic origin of the line. Morphologically, the cells were typically lymphoblastic. Cytochemically, they were slightly acid phosphatase-positive and negative for peroxidase, ASD chloroacetate esterase, and nonspecific esterase. Immunologically, they were found positive for common-ALL antigen (CALLA), Ia-associated antigen, terminal deoxynucleotidyl transferase (TdT), and cytoplasmic IgM (cIgM) and slightly positive for surface IgM (sIgM). Testing for Epstein-Barr virus (EBV) capsid antigen was also positive. Cytogenetic evaluations performed on initial, relapse, and cell line specimens each revealed the presence of a pseudodiploid cell line characterized by a consistent marker chromosome. GTG-, QFQ-, and RF-banding identified the marker as being derived from a translocation involving chromosomes #7 and #19: t(7;19) (q11;q13). Iso 7q, −5, −9, and +2 were also found in significant association with the marker and were viewed as demonstrating continued karyotypic evolution within the cell line. From these data, cell line #697 has been classified as a leukemic line of B-cell lineage in a transitional stage between pre-B and mature B cells.

Phenotypic characterization of cutaneous T-cell lymphoma. Use of monoclonal antibodies to compare with other malignant T cells.

We studied the surface-antigen pattern of T cells in peripheral blood and cell lines from patients with advanced cutaneous T-cell lymphoma (CTCL) or T-cell acute lymphoblastic leukemia. The antigen patterns of cutaneous T-cell lymphoma cells from peripheral blood and established cell lines were nearly identical; the cells were negative for human thymus antigen (OKT6 and NA1/34), positive for pan-T-cell (OKT3, 17F12, 10.2, and 9.6) and helper-T-cell-subset (OKT4) antigens, and negative for T-cell-subset antigens 3A1 and OKT8. In contrast, the phenotypes of malignant T cells from patients with acute lymphoblastic leukemia were heterogeneous, with at least five patterns of reactivity. The T-cell-specific antibody 3A1 was the only monoclonal reagent that clearly distinguished the peripheral-blood T cells in CTCL (3A1-) from those in acute lymphoblastic leukemia (3A1+). Moreover, 3A1 was the most reliable T-cell marker in acute lymphoblastic leukemia. We conclude that circulating CTCL (Sézary) T cells are homogeneous in their antigen phenotype and are derived from a well-differentiated 3A1-, OKT4+, OKT8- helper-T-cell subset.

Terminal deoxynucleotidyl transferase-positive acute myeloblastic leukemia.

Terminal deoxynucleotidyl transferase (TdT) is a biochemical marker for acute lymphoblastic leukemia (ALL). In studies of ALL at diagnosis, there are usually greater than 40% TdT-positive cells by indirect immunofluorescence, whereas acute myeloblastic leukemia (AML) shows less than 1% TdT-positive cells. Rare cases of TdT-positive AML have been reported. We present here three AML patients with TdT in 15%, 45%, and 90% of the leukemic blasts. The diagnosis of AML was established on the basis of morphology and cytochemistry, and the cases included one patient with Auer rods. Myeloperoxidase was present respectively in 20%, 90%, and 5% of the blasts. There was no Philadelphia chromosome present in the three cases. These results may indicate the simultaneous presence of lymphoid and myeloid populations, or the presence of a blast cell with both lymphoid and myeloid markers.

Subpopulations of human T lymphocytes. VI. Analysis of cell markers in acute lymphoblastic leukemia with special reference to Fc receptor expression on E-rosette-forming blasts.

A variety of surface markers, terminal deoxynucleotidyl transferase (TdT) activity, morphologic appearance, and cytochemical composition were studied in a group of 16 patients (13 children, 3 adults) with acute lymphoblastic leukemia (ALL). In 9 children, no surface markers were detected on lymphoblasts (null-type ALL). Leukemic blasts of 4 children formed E-rosettes. These E-rosette-forming blasts from 3 adult patients with ALL, were studied for the presence of Fc receptors. Of the leukemic blasts from these 7 patients, 2--76% expressed receptors for IgG Fc. Only 3 of 7 patients showed 9--42% receptors for IgM Fc. In addition, complement receptors were investigated in 6 of those 7 patients with T-cell ALL. Complement receptors were detected on 9--70% of the E rosette forming blasts from all 6 patients. TdT activity was elevated in T-cell ALL and in children with null-type ALL. The heterogeneity of Fc receptor expression on leukemic blasts in these patients demonstrates a malignant proliferation of T cells in different stages of differentiation or maturation. This observation might be helpful in subclassifying T-cell leukemias with regard to prognosis and the response to therapy.

Terminal transferase in acute lymphoblastic leukemia in gibbons

Terminal deoxynucleotidyl transferase, an enzyme which catalyzes the polymerization of deoxyribonucleoside triphosphates on a 3′-OH end of an initiator molecule in the absence of a template, has been suggested as a biological marker for human acute lymphoblastic leukemia. Examination of a cell line, 6G1, recently established from the peripheral blood of a gibbon ape with acute lymphoblastic leukemia showed the presence of terminal deoxynucleotidyl transferase. This enzyme after purification by successive column chromatography on DEAE-cellulose, phosphocellulose and hydroxyapatite, was found to have biochemical properties similar to those reported for terminal transferase from calf thymus and human leukemic cells. These studies suggest that terminal transferase can be used as a useful biological marker for acute lymphoblastic leukemia in both humans and subhuman primates.

Correlation of adenosine deaminase activity with cell surface markers in acute lymphoblastic leukemia.

Adenosine deaminase (ADA) activity has been measured in the lymphoblasts of 23 untreated patients with acute lymphoblastic leukemia and related to the presence or absence of immunologic cell surface markers. The mean ADA activity in the acute lymphoblastic leukemia population as a whole was increased fourfold over that in normal lymphocytes. 9 of the 23 patients were classified as thymus-derived (T-) cell acute lymphoblastic leukemia on the basis of erythrocyte rosette positivity; the remaining 14 patients had null-cell leukemia. The mean ADA activity (ADA U/mg protein) of T-cell lymphoblasts (102 U) was 3 times higher than the mean of null lymphoblasts (30 U). This difference is statistically significant (P less than 0.02). Measurement of ADA activity offers a biochemical method of distinguishing between immunological subtypes of lymphoblasts which may be of prognostic and therapeutic value.

Terminal transferase levels in chronic myelogenous leukemia in blast crisis and in remission

Terminal deoxynucleotidyl transferase (TdT) has been cited as a possible biological marker for acute lymphoblastic leukemia (ALL) and the blast phase of chronic myeloid leukemia (CML). In the present study high levels of TdT, comparable to levels present in normal thymus, were observed in circulating leukocytes of a CML patient in blast crisis. Sequential studies revealed disappearance of TdT from circulating leukocytes after remission was induced with vincristine and prednisone therapy. Blood TdT returned to moderately elevated levels in the absence of detectable circulating blasts, 5 months before the reappearance of blast crisis. During blast crisis the malignant cells were devoid of myeloid features on light and electron microscopy and were peroxidase negative and periodic acid-Schiff (PAS) positive. These studies demonstrate that elevated TdT levels may occur with or without detectable morphologic evidence of circulating blast cells. The presence of complement receptors on agranular blasts during the blast phase of chronic myeloid leukemia is a new finding.

Lymphocyte Surface Markers in Acute Lymphoblastic Leukemia of Adults

Leukemic cells from eight adult patients with acute lymphoblastic leukemia (ALL) were examined for the T-lymphocyte (T-cell) marker of sheep erythrocyte (E) receptors and for B-lymphocyte (B-cell) markers of surface immunoglobulins (SIg), complement receptors and FC receptors. Six of these patients' leukemic cells were devoid of both T- and B-cell markers, and therefore were "null" cells. The leukemic lymphoblasts of a 79-year-old patient had all B-cell markers, including monoclonal IgG K and receptors for complement and Fc. In cells from another patient (21 years old), only complement receptors were found. A review of the literature revealed that, similar to childhood ALL, adult cases of ALL were mostly of "null" cell type. All of the T-cell type was found in approximately 20% of patients. ALL of the B-cell type occurred only rarely. The latter cell type appeared to occur mainly in the middle-aged and the elderly.