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   2019| October-December  | Volume 44 | Issue 4  
    Online since July 20, 2020

 
 
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ORIGINAL ARTICLES
Study of Wilms’ tumor 1 gene expression in patients with acute myeloid leukemia
Ebtesam I Ahmad, Ghada M El-Akad, Weaam I Ismail, Ahmed A.R Al Nagar
October-December 2019, 44(4):195-203
DOI:10.4103/ejh.ejh_26_19  
Background The Wilms’ tumor 1 (WT1) gene is overexpressed in patients with acute leukemias. Many studies have reported the importance of quantitative assessment of WT1 expression as a molecular marker of minimal residual disease monitoring. However, biological significance and the prognostic effect of WT1 overexpression in acute myeloid leukemia (AML) remain elusive. Subjects and methods A total of 42 patients with newly diagnosed AML were included in the study. Also, 42 individuals matched for age and sex were enrolled as controls. Immunophenotyping, cytogenetic analysis, and quantitative assessment of WT1 gene transcripts were performed using real-time PCR. Results WT1 overexpression was detected in 73.8% of our patient group. There was a statistically significant decrease in the probability of achieving complete remission with shorter overall survival and event-free survival in the WT1 overexpression group compared with the downexpression group (P=0.035, 0.045, 0.011, respectively). Application of multivariate analysis showed that WT1 overexpression by itself is an independent and negative indicator for predicting overall survival and disease-free survival of AML patients. Conclusion WT1 overexpression is an independent negative prognostic marker that could be used to evaluate response to induction chemotherapy and prognosis of AML patients.
  1,047 124 1
Leukemia stem cell markers (CD 123 and CD25) are poor prognostic markers in patients with adult acute myeloid leukemia
Botheina A.T Farweez, Azza E Hashem, Walaa A Elsalakawy
October-December 2019, 44(4):208-217
DOI:10.4103/ejh.ejh_35_19  
Background Acute myeloid leukemia (AML) still remains a challenge for hematologists. Though an impressive number of prognostic factors have been identified in AML, it still ranks as one of the highest cancer-related deaths. Several studies have indicated its origin from a rare population of leukemic cells, known as leukemic stem cells, which initiate the disease and contribute to frequent relapses. Leucocyte interleukin-3 receptor α (CD123) and leukocyte interleukin-2 receptor α (CD25) are regarded as markers of leukemia stem cells. Aim The aim of this study was to investigate CD123 and CD25 expressions in newly diagnosed patients with AML by flow cytometry and correlate their expression with disease prognostic parameters and patients’ outcome at day 28 of therapy. Patients and methods This study was conducted on 30 newly diagnosed patients with AML admitted to Ain Shams University Hospitals. The expression of CD123 and CD25 was assessed, where gated blast cells were stained for CD45, CD38, CD34, CD123, and CD25. Results In the current study, CD123 was expressed in 13/30 (43.3%) patients, and CD25 was expressed in 4/30 (13.3%) patients. CD123 expression positively correlated with higher total leukocytic count and bone marrow blast percentage and CD25 expression. Both CD123 and CD25 expression had a significantly poor effect on outcome even in the good prognostic cytogenetic subgroups. Conclusion Results of our study clearly demonstrate the poor prognostic significance of CD123 and CD25 expression in patients with AML. This may represent an additional prognostic tool in risk-stratified management of patients with AML.
  896 91 -
Reduced serum vitamin D levels in Egyptian adults with chronic primary immune thrombocytopenia (responders and nonresponders): an Egyptian single-center study
Walaa A Esalakawy, Abdel R.A Soliman, Sana A Mohammed, Alia M Saeed
October-December 2019, 44(4):246-253
DOI:10.4103/ejh.ejh_34_19  
Introduction Immune thrombocytopenia (ITP) is a disorder characterized by immune-mediated accelerated platelet destruction and suppressed platelet production. Low vitamin D levels have been found in several autoimmune diseases, such as rheumatoid arthritis, systemic lupus erythematosus, systemic sclerosis, type 1 diabetes mellitus, multiple sclerosis, inflammatory bowel diseases, and autoimmune gastritis. The mechanisms underlying the link between vitamin D and autoimmunity are not completely understood but probably are associated with its anti-inflammatory and immunomodulatory functions. Patients and methods The study included 80 adult participants, comprising 40 patients with primary ITP (they were divided into 20 responders and 20 nonresponders), 20 cases of thrombocytopenia owing to non-ITP causes (eight acute myeloid leukemia, five myelodysplastic syndrome, and seven with aplastic anemia), and 20 healthy controls. All were subjected to measurement of serum 25-monohydroxyvitamin D level with ELISA. Results We found that vitamin D levels were significantly lower in patients with ITP (range=2–40 ng/ml; mean±SD=17.29±10.96 ng/ml) and thrombocytopenia owing to non-ITP causes (range=10–40 ng/ml; mean±SD=21.05±8.31 ng/ml) in comparison with normal healthy controls (range=10–65 ng/ml; mean±SD=36.70±16.30 ng/ml) (P=0.000), but there was no statistically significant difference between levels in ITP vs non-ITP thrombocytopenia (P=0.225). When comparing vitamin D levels in patients with ITP in relation to response to first-line treatment with corticosteroids, there was no statistical significant difference regarding vitamin D levels (mean±SD=15.73±10.00 ng/ml in responders and mean±SD=18.85±11.89 ng/ml in nonresponders), with a P value of 0.374. Conclusion Vitamin D levels are lower among patients with ITP in relation to healthy controls. Vitamin D may play a role in the pathogenesis of ITP.
  776 67 -
Sensitivity of flow cytometry immunophenotyping compared with bone marrow morphology in diagnosis of multiple myeloma
Eman A Yousof, Eman H Ahmed, Shima G Mansor, Hanan O Mohamed, Ahmad F Thabet, Douaa M Sayed
October-December 2019, 44(4):227-231
DOI:10.4103/ejh.ejh_42_19  
Background The diagnosis of multiple myeloma (MM) is challenging, as it is based on several factors, including serum and urine protein electrophoresis, immunofixation, bone marrow (BM) aspirate, and biopsy, in addition to immunophenotyping. Moreover, quantification of BM plasma cells (PCs) by conventional morphology is a mandatory test for the diagnosis and response assessment in MM. One alternative would be to use multiparameter flow cytometry immunophenotyping to quantify BMPCs, although this is currently still limited to research studies and the differential diagnosis of unusual cases. Aim The aim was to determine the diagnostic sensitivity of flow cytometry in the detection of abnormal PCs in BM specimens of MM compared with BM morphology. Patients and methods We retrospectively analyzed bone marrow aspiration (BMA), bone marrow biopsy (BMB), and immunophenotypic reports of 50 newly diagnosed patients with MM. Patients’ data were collected and analyzed. Results BMA of 43 (86%) patients showed PCs greater than or equal to 10%; BMB of 35 of them showed PCs greater than or equal to 10%, and five patients were with poor BMB quality. The sensitivity of BMB in detection of PCs was 90% and 86% for BMA. Concerning immunophenotyping, the gate of mononuclear cells was in the range of 4–88%; those gated cells were 100% positive for CD138 and CD38. CD56 was positive in 84% whereas CD19 was negative in 96%. Overall, 46% were κ restricted and 54% were λ restricted, with 100% sensitivity to malignant PCs. Conclusion Unlike morphologic and histologic studies, FC is not dependable for providing quantitative information about MM but provides qualitative information for assessing the immunophenotype and light chain clonality of PCs.
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Association of the cyclin D1 G870A polymorphism with risk of multiple myeloma in a group of Egyptian patients
Dina El Dahshan, Raafat AbdelFattah
October-December 2019, 44(4):204-207
DOI:10.4103/ejh.ejh_18_19  
Background Multiple myeloma (MM) is a clonal bone marrow disease characterized by the neoplastic transformation of mature B cells. Single-nucleotide polymorphisms in the Cyclin d1 CCND1 gene were reported with an increased susceptibility to various human cancers such as breast cancer, bladder cancer, colorectal cancer, and lung cancer. Aim The aim of this study was to detect the distribution of CCND1 G870A genotypes in a group of MM patients and in controls in an attempt to reveal the possible association between the presence of the polymorphism (G870A) and the risk of MM in an Egyptian population. Patients and methods The PCR-restriction fragment length polymorphism technique was used for the detection of CCND1 G870A polymorphism in 56 MM patients and 70 healthy individuals as a control group. Results The homozygous GG genotype was present in 10.7% of patients, heterozygous AG in 48.2%, and homozygous AA in 41.1%. Carriers of CCND1 G870A polymorphism did not show increased risk to MM [AG genotype odds ratio (OR)=0.928, P=0.899 and GG genotype OR=1.295, P=0.499]. Patients aged at least 60 years had a higher frequency of mutant CCND1 than controls (88.2 and 64.3%), but this difference did not reach statistical significance (OR=4.167, P=0.128). Conclusion Carriers of the pleomorphic genotype of CCND1 G870A did not show increased susceptibility to MM. Larger population-based studies are needed to validate the association of CCND1 G870A polymorphism with MM.
  604 86 -
Leukocyte-associated immunoglobulin-like receptor-1, T-cell leukemia/lymphoma protein-1, and nuclear factor κB expression in childhood acute lymphoblastic leukemia
Mona H.Y Alrayes, Reham H.M Hammad, Botheina A.T Farweez, Nayera H.K Elsherif, Doaa A.A Aly
October-December 2019, 44(4):218-226
DOI:10.4103/ejh.ejh_5_19  
Background Immune regulation is crucial for the pathogenesis of childhood acute lymphoblastic leukemia (ALL). Leukocyte-associated immunoglobulin-like receptor-1 (LAIR-1) is an immune regulator, expressed by T and B immune cells. T-cell leukemia/lymphoma protein-1 (TCL1) is a coactivator of α-serine/threonine-protein kinase, and when dysregulated, it causes lymphomagenesis and cancer progression. Nuclear factor-κB (NF-κB) is a transcription factor that regulates genes involved in immune responses. This study aimed to study the expression of LAIR-1, TCL1, and NF-κB on blasts in childhood ALL with evaluation of their prognostic value. Patients and methods This descriptive cross-sectional study was conducted on 60 newly diagnosed childhood ALL cases. They were divided into group 1 (n=47) of B-cell ALL and group 2 (n=13) of T-cell ALL. The expression pattern of LAIR-1, TCL1, and NF-κB on blasts was assessed using flow cytometry. Results Correlation studies in total patients with ALL (n=60) revealed significant positive correlation between percentage of blasts expressing LAIR-1% and total leukocytic count (r=0.262, P=0.043) and percentage of blasts infiltrating the peripheral blood (r=0.292, P=0.025). A significant positive correlation of LAIR-1 mean fluorescence intensity with percentage of blasts expressing CD13 (r=0.293, P=0.026) and CD33 (r=0.373, P=0.004) was found. In group 2, a significant positive correlation was seen between percentage of blasts expressing TCL1 and LAIR-1 mean fluorescence intensity (r=0.566, P=0.044). According to Cox regression analysis, the percentage of blasts expressing NF-κB was seen to significantly increase the risk of death, in childhood ALL (hazard ratio=1.085, 95% confidence interval=1.02–1.16, P=0.01). Conclusion LAIR-1 expression carries the potentiality of being a bad prognostic marker in childhood ALL. A relation is suggested between the expression of both LAIR-1 and TCL1 on blasts. NF-κB expression might negatively influence the survival rate in childhood ALL.
  581 70 -
Coexistence of (CCND1/PRAD-1/BCL1) juxtaposition and drug resistance gene is a marker for vincristine-resistant B-non-Hodgkin’s lymphoma
Sahar El Gammal, Eman Mosad
October-December 2019, 44(4):254-259
DOI:10.4103/ejh.ejh_27_19  
Background CCND1/PRAD-1/BCL1 juxtaposition activates immunoglobulin heavy chain (IgH) enhancing effect on cyclin D1 expression. This leads to overexpression of cyclin D1, a cell-cycle regulator. Cyclin D1 can function as an oncogene growth advantage for tumor cells by way of cell-cycle progression. Multidrug resistance (MDR)1 gene expression leads to overproduction of P-glycoprotein (P-gp) which leads to chemotherapy resistance causing poor prognosis and short survival [overall survival (OS) and disease free survival(DFS)] in B-non-Hodgkin’s lymphoma (NHL). Objective To evaluate BCL1/IgH juxtaposition by fluorescent in-situ hybridization (FISH) and MDR1 protein expression by immunohistochemistry in B-non Hodgkin lymphoma (B-NHL) and to clarify if these genes have some relevance to developing vincristine treatment resistance of B-NHL and their correlation with clinical outcome. Patients and methods Fifty-three bone marrow biopsy samples with B-NHL were included in the study for the expression of t(11;14) BCL1/IgH in various B-cell lymphomas by FISH and MDR1/P-gp by immunohistochemistry and its relation to the outcome and chemotherapy resistance. Results In this study t(11;14) detected by FISH was positive in 64.2%. It was positive in (66.7%) of mantle cell lymphoma, in 64.3% of small cell lymphoma, and in 60% of large cell lymphoma. The detection frequency of MDR1/P-gp by immunohistochemistry in mantle cell lymphoma, small cell lymphoma and large cell lymphoma, is 80, 82, and 80%, respectively. Conclusion In this study, the frequency of t(11;14) detection by FISH and MDR1/P-gp expression by immunohistochemistry in Egyptian patients were consistent with other studies and the coexistence of MDR1/P-gp and t(11;14) in B-NHL is important before treatment to allow the design of novel drug regimens.
  546 57 -
Positive expression of FOXP1 protein as a good prognostic factor in childhood acute lymphoblastic leukemia: a retrospective study
Shaaban Redwan Helal, Eman Mosad, Mohamed Zakaria Abd-Elrahman, Sahar ElGammal, Marwa Saad Mohamed
October-December 2019, 44(4):237-245
DOI:10.4103/ejh.ejh_37_19  
Background Deficiency of FOXP1 in early B-lymphoid precursors results in a block at the transition of pro-B to pre-B-cell stage and severely reduces the peripheral mature B-cell compartment. Objective The objective of the current study was to detect the prognostic value of positive FOXP1 protein expression on acute lymphoblastic leukemia (ALL) in children and adults evaluated by disease-free survival (DFS) and overall survival and its relation with BCR-ABL fusion gene in those patient. Materials and methods A retrospective study was carried out on 50 bone marrow samples from fixed bone marrow aspirates taken from patients previously diagnosed with ALL at South Egypt Cancer Institute. They were classified into 25 BCR-ABL-positive samples and 25 BCR-ABL-negative samples. Results Positive expression of FOXP1 protein in B lymphocytes in patients with ALL was associated with better overall survival and DFS, especially in children. Coexistence of both FOXP1 and BCR-ABL proteins in the patients with ALL was associated with good prognosis than BCR-ABL-positive patients with negative expression of FOXP1. Conclusion The positive expression of FOXP1 protein in B lymphocytes in pediatric patients with ALL and its association with better overall survival and DFS indicate that FOXP1 protein expression in pediatric ALL is a good prognostic marker. Coexistence of both FOXP1 and BCR-ABL proteins in the patients with ALL appeared as a good prognostic factor.
  534 60 -
Impact of cytomegalovirus viremia on allogeneic peripheral blood stem cell transplantation in patients with acute leukemia: a single Egyptian center experience
Mohammed O Azazzi, Raafat A Soliman, Gehad A.M Hasan, Mary G Naguib
October-December 2019, 44(4):232-236
DOI:10.4103/ejh.ejh_31_19  
Background Hematopoietic stem cell transplantation is now established as a standard therapeutic modality for a variety of malignant and nonmalignant diseases. The first successful allogeneic hematopoietic stem cell transplantation was done with bone marrow as the source of hematopoietic stem cells in 1968. Aim This retrospective study evaluates the impact of cytomegalovirus (CMV) viremia in patients with acute myeloid leukemia and acute lymphoblastic leukemia after allogeneic stem cell transplantation. Patients and methods Patients with acute leukemia (AML and ALL) who underwent allogeneic peripheral blood stem cell transplantation at Nasser Institute, in the time period between 2015 and 2016 (2 years) were included in the study. Results There were 112 patients: 69 men (61.6%) and 43 women (38.4%). Age: mean=30.1±12.04, range 4–58. Overall survival of the CMV positive group is 60% which is lower than that in the CMV negative group which is 69.1% with a statistically insignificant P value of 0.3. Overall survival of CMV positive and acute graft-vs-host disease (aGVHD) patients is 53.8 which is lower than that of CMV negative patients which is 69.6% but it is statistically insignificant (P=0.1). This may be attributed to non-CMV–non-aGVHD-related mortality in acute leukemia (as disease relapse). Conclusion The use of CMV prophylaxis routinely with new agents with lower toxicity, especially in patients at high risk of CMV replication, might reduce the incidence of CMV replications, reducing the morbidity and mortality, and according to some studies may also reduce the incidence of aGVHD and its related morbidity and mortality.
  499 59 -
ERRATUM
Erratum: Prognostic significance of progranulin and B7-H4 in patients with diffuse large B-cell lymphoma

October-December 2019, 44(4):260-261
DOI:10.4103/1110-1067.290226  
  365 48 -