Nadia El Menshawy

Antiplatelet antibodies contribute to thrombocytopenia associated with chronic hepatitis C virus infection

Abstract Thrombocytopenia is one of the most frequent hematological manifestations of hepatitis Cvirus (HCV) infection; which typically worsens with progression of the liver disease and can become a major clinical complication. Several mechanisms have been postulated to explain thrombocytopenia in HCV hepatic patients, including immune mechanisms. The aim of the present work is to investigate the role of immune mechanisms as a causative agent of thrombocytopenia in HCV hepatic patients. The study included 50 hepatic patients with HCV infection (30 with thrombocytopenia and 20 with normal platelets counts). Platelets associated glycoprotein specific antibodies were evaluated by flow cytometry and confirmed by quantitative monoclonal immobilization of platelet antibodies (MAIPA). The frequency of platelet associated immunoglobulin (PAIg) in thrombocytopenic HCV positive hepatic patients by FCM was 86·7, 83·3, 46·7 and 33·3% for total PAIg, PAIgG, PAIgM and PAIgA respectively. MAIPA found platelet specific antibodies in 26/30 (86·7%) of patients. The most likely target antigen for platelets antibodies were glycoprotein (GP) IIb/IIIa (30%), followed by GP IIIa (20·5), GP IIb (13·3%), GPIb (13·3%), then GPIa (10%). The platelets count was inversely correlated to the levels of platelets GP specific antibodies (r=−0·42, p=0·024), and significantly parallel to spleen size (p=0·024). Platelet associated glycoprotein specific antibodies represent a common mechanism inducing thrombocytopenia in patients with chronic HCV infections.

Clinicopathologic Effect of DNMT3A Mutation in Adult T-Cell Acute Lymphoblastic Leukemia

BACKGROUND The present study aimed to determine the frequencies and clinicopathologic effect of a DNMT3A [DNA (cytosine-5)-methyltransferase 3A] mutation in patients with adult T-cell acute lymphoblastic leukemia (T-ALL). PATIENTS AND METHODS A total of 64 patients with T-ALL who had been admitted to Mansoura University Oncology Center were included in the present study. For all patients, DNA extraction and amplification with sequencing analysis using the 310 ABI genetic analyzer for detection of a mutation (R882H). RESULTS The DNMT3A mutation (R882H) was found in 12 of the 64 patients (18.8%). The DNMT3A mutation was frequently detected in the older age group and was associated with high leukocytic counts, a high bone marrow blast cell percentage, and the frequent presence of extramedullary disease. However, it was not associated with the hemoglobin level, red blood cell count, or platelet count. The patients with mutant T-ALL had a low tendency to achieve remission after induction. These patients had significantly shorter overall survival and shorter disease-free survival compared with those with wild-type T-ALL (P = .037 and P = .006, respectively). CONCLUSION DNMT3A is frequently mutated in T-ALL and is associated with distinct clinicopathologic entities and a poor prognosis. These findings could help in risk stratification and treatment decisions for patients with T-ALL.

Effect of splenectomy on platelets associated antibodies in hepatitis C patients with thrombocytopenia

Abstract Background This study aimed to assess the role of splenomegaly as a source of platelet-associated immunoglobulins (PAIgs) in thrombocytopenic patients with chronic hepatitis C virus (HCV) infection. Subjects and methods The present study was conducted on 63 subjects categorized as follows. Group 1: Included 63 cases diagnosed as patients with HCV liver cirrhosis combined with thrombocytopenia before splenectomy. Group 2: Included the same 63 cases one week after splenectomy. For all subjects included in this study platelets counts were evaluated as well as PAIgs (total Igs, IgG, IgM, and IgA). Results All patients were thrombocytopenic before undergoing splenectomy (platelet counts [median 68.5; range 44–95]). After splenectomy all patients achieved normal platelet counts (median 180; range 108–235). The mean ± SD of PAIgs was 64.2 ± 9.6 for total IgG, 53.6 ± 8.1 for IgG, 3.8 ± 2.1 for IgM, 6.7 ± 4.7 for IgA presplenectomy versus postsplenectomy 13.4 ± 19.3 for total Igs, 5.4 ± 1.8 for IgG, 1.9 ± 1.06 for IgM, 2.1 ± 0.9 for IgA, and the differences between pre and postsplenectomy figures were statistically significant (P < 0.001). The correlation studies between platelet count and PAIgs level in patients with chronic HCV infection presplenectomy revealed that there is significant negative correlation between platelet counts and all immunoglobulins (total Igs: r − 0.804, P = 0.000; IgG: r − 0.907, P = 0.000; IgM: r − 0.467, P = 0.002; and IgA: r − 0.519, P = 0.000). Conclusion Autoimmune mechanism plays an important role in the HCV-associated thrombocytopenia and spleen is a major source of PAIgs.

Soluble angiopoietin-2/sTie2 receptor ratio is an independent prognostic marker in adult acute myeloid leukemia.

AimAngiogenesis is the formation of new blood vessels from preexisting one. The angiopoietins act a central role in these process. The aim of the present study is to the assess the impact of the circulating levels of angiopoietin-1 (Angi-1), Angiopoietin-2 (Angi-2), soluble Tie2 receptor (sTie2), and calculated Angi-2/sTie2 ratio on overall survival in 71 acute myeloid leukemia patients and 19 normal controls.Materials and methodsAng-2, and calculated angi-2/sTie values were significantly higher in AML patients as compared to healthy volunteer (P= 0.002, 0.015 respectively) on the other hand Angi-1 was not significantly different in patients and control.ResultsIn univariate Cox proportional hazards model Angi-2, sTie2, angi-2/sTie2 ratio were predictive of poor survival. In multivariate analysis the calculated angi-2/sTie2 ratio is independent prognostic factor, with relative risk of 3.939, with 95% confidence interval 0.002–0.001.ConclusionThe calculated angiopoietin-2/sTie2 ratio represent an independent prognostic factor in AML and its value should be considered when considering anti angiogenic therapy.

Wilms tumor 1 gene mutations in patients with cytogenetically normal acute myeloid leukemia.

Objective: This study aimed to assess the prognostic impact of Wilms tumor 1 (WT1) mutations in cytogenetically normal acute myeloid leukemia (CN-AML) among Egyptian patients. Materials and Methods: Exons 1, 2, 3, 7, 8, and 9 of WT1 were screened for mutations in samples from 82 CN-AML patients out of 203 newly diagnosed AML patients, of age ranging from 21 to 74 years, using high-resolution capillary electrophoresis. Results: Eleven patients out of 82 (13.41%) harbored WT1 mutations. Mutations were detected in exon 7 (n=7), exon 9 (n=2), exon 8 (n=1), and exon 3 (n=1), but not in exons 1 or 2. There was no statistically significant difference between the WT1 mutants and wild types as regards age, sex, French-American-British subtypes, and the prevalence of success of induction remission therapy (p=0.966; 28.6% vs. 29.3%). Patients with WT1 mutations had overall survival lower than patients with the wild type (HR=1.38; 95% CI 4.79-6.86; p=0.004). Conclusion: CN-AML patients with WT1 mutations have poor clinical outcome. We recommend molecular testing for WT1 mutations in patients with CN-AML at diagnosis in order to improve risk stratification of those patients.

Prognostic relevance of CD69 expression in B cell chronic lymphocytic leukemia

This study aimed to determine the prognostic impact of CD69 expression in B cell chronic lymphocytic leukemia (CLL). The study included 153 B cell chronic lymphocytic leukemia (B-CLL) recruited to Mansoura Oncology Center. Cellular CD69 expression was determined by multicolor flow cytometry in parallel with PCR amplification and sequencing analysis to mutational status of immunoglobulin variable heavy chain (IgVH). The number of CLL patients positively expressed CD69 was 47 out of 153 (30.7%). There is significant association between CD69-positive expression and unmutated IgVH and between CD69 negative expressions with mutated IgVH gene (P < 0.007). The patients positively expressed CD69 were younger in age and had higher leukocyte count, advanced RIA stages III and IV, and high β2 microglobulin as compared to negative ones. CLL patients group positively expressed CD69 had short overall survival and progression disease-free survival (PDFS) as compared to negative ones. Cox proportional hazard regression model showed that positive CD69 expression is an independent prognostic biomarker for progression-free survival (HR for CD69 vs IgVH was 1.7 vs 1.6, p = 0.04 vs 0.02) and OS (HR 1.8 vs 1.9, p = 0.03 vs 0.01). In conclusion, positive CD69 expression could predict adverse outcome; it could be included in scoring system of CLL as simple, easy applicable to stratify early progressive patients and enabling timely therapeutic decisions to improve outcome. Also, it could be a target for therapy for B cell CLL.

CD58; leucocyte function adhesion-3 (LFA-3) could be used as a differentiating marker between immune and non-immune thyroid disorders

The link between Graves’ disease (GD) and Hashimoto’s thyroiditis (HT) has been debated for decades due to the shared pathological and immunological components. Immune intolerance and inappropriate immune reaction against self-thyroid cells are distinctive features of both diseases, but definitive data for the clinical presentation of autoimmune thyroid disease remains unclear. To analyse the expression of T-regulatory cells, CD58, the CD4/CD8 ratio and the neutrophil/lymphocyte ratio and to determine if these parameters could be used as differentiating markers between auto- and non-immune thyroid diseases, 75 patients were enrolled in this study—40 with autoimmune thyroid disease (HT and GD ), 15 with non-immune thyroid disease, and 20 healthy controls. Multicolour flow cytometry was used to analyse CD58, T-regulatory cells (Treg) expressing CD4, CD25, HLA-DR and CD8 using different stained fluorescent labelled monoclonal antibodies. The neutrophils and lymphocyte ratio was also measured. Lower expression of Treg with higher expression of CD58 (LFA-3) was found in the autoimmune diseases when compared with the non-immune and control groups. ROC analysis showed that CD58 with sensitivity 88% and specificity 100% with cut-off value more than or equal to 29.9 indicates Hashimoto’s disease, while lower value indicates colloid goitre, and higher or equal to 29.84 indicates Graves’ disease and lower indicates colloid goitre with 100% sensitivity and specificity. CD58 could be used as differentiating marker between immune and non-immune thyroid disorders.

Regulatory T cells in chronic lymphocytic leukemia

Controversial results concerning the prognostic value of the regulatory T (Treg) cell percentage in peripheral blood of chronic lymphocytic leukemia (CLL) patients have been reported in the previous studies. This study aimed to estimate the prognostic relevance of Treg cells in untreated CLL patients at diagnosis. CLL patients showed significantly higher Treg cell percentage and Treg cell counts as compared to that identified in healthy normal controls (P < 0.01 for both). Furthermore, CLL patients with high LDH, β2 microglobulin levels, and those positive for CD38% express significantly higher Treg cell percentage as compared to those patients with normal LDH, β2 microglobulin levels, and negative CD38%. Also, the percentage of Treg cells was significantly higher among CLL patients having autoimmune cytopenias. In conclusion, Treg cell percentage is higher in CLL patients as compared to normal healthy controls and related to advanced stages as well as poor prognostic markers. Treg cell manipulation may represent a future strategy for management of CLL patients.