ABSTRACT
@#Introduction: Chronic lymphocytic leukaemia (CLL) is the most frequent adult leukaemia in the Western world. The clinical presentation varies greatly, from very indolent cases to those with aggressive and fast advancing disease. This variation has significant implications for clinical approaches, therapeutic tactics, and, ultimately, survival durations from diagnosis. Acquired chromosomal aberrations play a key role in CLL aetiology. Due to difficulty to obtain abnormal metaphases for analysis, few methods such as fluorescence in-situ hybridization (FISH) and multiplex ligation-dependent probe assay (MLPA) were employed to detect chromosomal aberration however the methods are limited to specific locus only. Thus, this study is aimed to detect the chromosomal aberrations using DNA microarray platform. Methods: In this retrospective study, DNA archive obtained from 7 CLL patients which collected at diagnosis and subjected to Affymetrix CytoScan® 750K single nucleotide polymorphism (SNP) array following the manufacture procedure. The raw data obtained were analysed using the Chromosome Analysis Suite (ChAS) software (Affymetrix) using annotations of genome version GRCh38 (hg38). Result: Out of 7 patients, 4 of them showing deletion of 13q while 3 of them showing deletion of 14q in various region . Some of the deleted loci were too small (0.42-0.6Mb) to be detected by conventional cytogenetic analysis (CCA). There was also the presence of additional chromosomal aberrations that could be missed by CCA, FISH, or MLPA due to cryptic deletion or duplication that was as small as 0.4MB in size. Conclusion: The present study showed that low resolution chromosomal aberration was able to be detected using DNA microarray platform in comparison to CCA, FISH and MLPA.
ABSTRACT
Chronic lymphocytic leukemia is a neoplastic entity pertaining to lymphocytes when they get accumulated in either lymph nodes thereby call as Small lymphocytic lymphoma or spilled up in the circulation of the blood. These lymphocytes are mature looking, relatively immunologically competent usually expressing B cell phenotype markers. The median age of affection of patients at the time of diagnosis is 71 years and its incidence increases with age. Its incidence in an age group <50 years is quite uncommon accounting only to 10-15% of total diagnosed cases. Here we are presenting such a rare case report of chronic lymphocytic leukaemia where age of affection is less than 40 years
ABSTRACT
BACKGROUND: Chronic lymphocytic leukaemia (CLL) is a neoplasm of B-cells characterized by variable prognosis. Exploring the proteome of CLL cells may provide insights into the disease. Therefore, eleven proteomics experiments were conducted on eleven primary CLL samples. RESULTS: We reported a CLL proteome consisting of 919 proteins (false discovery rate (FDR) 1%) whose identification was based on the sequencing of two or more distinct peptides (FDR of peptide sequencing 1%). Mass spectrometry-based protein identification was validated for four different proteins using Western blotting and specific antibodies in different CLL samples. Small sizes of nucleolin (~57 kDa and ~68 kDa) showed a potential association with good prognosis CLL cells (n = 8, p < 0.01). Compared with normal B-cells, CLL cells over-expressed thyroid hormone receptor-associated protein 3 (THRAP3; n = 9; p = 0.00007), which is implicated in cell proliferation; and heterochromatin protein 1-binding protein 3 (HP1BP3; n = 10; p = 0.0002), which promotes cell survival and tumourogenesis. A smaller form of HP1BP3, which may correspond to HP1BP3 isoform-2, was specifically identified in normal B-cells (n = 10; p = 0.0001). HP1BP3 and THRAP3 predicted poor prognosis of CLL (p 0.05). Consistently, THRAP3 and HP1BP3 were found to be associated with cancer-related pathways (p 0.05). CONCLUSIONS: Our findings add to the known proteome of CLL and confirm the prognostic importance of two novel cancer-associated proteins in this disease.
Subject(s)
Leukemia, Lymphocytic, Chronic, B-Cell , Biomarkers, Tumor/analysis , Mass Spectrometry , Transcription Factors/analysis , Nuclear Proteins/analysis , Blotting, Western , Chromatography, Liquid , Proteomics , DNA-Binding Proteins/analysisABSTRACT
@#Introduction: Chronic Lymphocytic Leukaemia (CLL) is a common type of leukaemia in persons of predominantly European descent but is rare in the Asian population. Disparities in CLL incidence among people of Asian and European descent may be related to the genetic make-up of the two different populations. Hypermethylation event might be one of the silencing mechanisms that inactivate the tumour suppressor genes in CLL. The aim of this study was to determine the hypermethylation status of p16INK4a and p15INK4b among CLL patients and normal individuals. Materials & Methods: A total of 25 CLL patients and 25 normal individuals were recruited for this study and their genomic DNA were extracted from the peripheral blood. The hypermethylation status of p16INK4a and p15INK4b were determined using Methylation Specific-PCR (MS-PCR) whereas DNA sequencing method was applied to selected samples for validation of the MS-PCR results. We also evaluated the association between hypermethylation of these genes with the clinical and demographic characteristics of each group of subjects. Results: Among the CLL patients, p15INK4b partialmethylation occurred in 6 (24%) subjects while methylation occurred in 1 (4%) subject. All the remaining patients were unmethylated at p15INK4b. All the samples showed unmethylation at p16INK4a. Statistically significant associations were found between p15INK4b hypermethylation with the presence of CLL (p=0.01) and with race (p=0.02). Conclusion: Further study using a larger sample size is warranted to explore the significance of DNA methylation incidence among the CLL patients of the Malaysian population. Hence, we suggest that hypermethylation at p15INK4b has a huge influence that kick-starts CLL disease among Malaysians and MS-PCR technique is applicable to be used in methylation study.
ABSTRACT
@#Isolated third nerve palsy as the sole manifestation of meningitis is rare. We describe a patient with chronic lymphocytic leukemia who developed third nerve palsy due to HSV2 meningitis. HSV2 PCR was positive in CSF and the patient was partially recovered upon treatment with acyclovir
ABSTRACT
Objectives: Chronic lymphocytic leukaemia (CLL) is associated with abnormalities of the B-Cell Receptor (BCR) signalling, including low responsiveness to antigenic stimulation and constitutive phosphorylation of several components of the signalling pathway. In Bcells, BCR-mediated signalling is regulated in part by the amount of membrane cholesterol. It was observed that Statins, pharmacological inhibitors of cholesterol synthesis, induce apoptosis of CLL cells in vitro and in vivo. Having previously reported that ectopic expression of CD5 in a B-cell line stimulated the transcription of genes involved in the synthesis of cholesterol, we investigated the expression and synthesis of cholesterol in CLL B-cells. Study Design & Methodology: Plasma membrane cholesterol in CLL cells was evaluated by staining with Filipin and Flow cytometry in 26 patients. CLL cells were cultured with Lovastatin and subG1 cells and Gumprecht’s shadows counted thereafter; surface expression of IgM, CD19 and CD5 was analysed. The expression of cholesterol synthesis genes was investigated in transcriptomic data from the MILE project (150 CLL and 110 controls). Results: We confirmed that leukemic B-cells contained more cholesterol in their plasma membranes than their normal counterparts. An enhanced expression of genes involved in the synthesis of cholesterol in CLL as compared to healthy controls was observed. Interestingly, among the 150 CLL patients analyzed, four cholesterol synthesis genes were activated in 65 “Ig-mutated” (M) in comparison to 69 “Ig-unmutated” (UM) CLLs. Leukemic cells cultured with Lovastatin exhibited a dose-response apoptosis, however surface IgM expression was unaffected and CD19 and CD5 were downregulated at highest concentrations only. Conclusions: High membrane cholesterol in CLL cells may explain their sensitivity to Statins, with a potential difference between UM- and M-CLL.
ABSTRACT
Verbena officinalis L., Verbenaceae, commonly known as vervain, is a plant widely used in medicine. Despite of its widespread use in different traditional practices, the mechanisms of pharmacological actions of the plant and its volatile oil are still unclear. We evaluated the pro-apoptotic activity of V. officinalis essential oil and of its main component, citral, on lymphocytes collected from ten patients with chronic lymphocytic leukaemia (CLL), a disease in which a faulty apoptotic mechanism is still retained one of the primary pathogenic events, by adding to treated mononuclear cells, annexin-V, propidium iodide, and CD19. Apoptosis was also evaluated using anti-active-caspase-3 monoclonal antibody after permeabilization of the cells. Both V. officinalis essential oil and citral were found able to induce apoptosis in CLL cells and to activate caspase-3, which is considered the way by means they active apoptosis in B neoplastic cells. This data further support evidences that indicate natural compounds as possible lead structure to develop new therapeutic agents for CLL.
ABSTRACT
Chronic lymphocytic leukaemia (CLL) is a haematological malignancy for which reliable prognostic markers are needed in view of its clinical heterogeneity. In approximately 50 percent of CLL patients, immunoglobulin (Ig) rearrangements are modified by somatic hypermutation (SHM), a process that represents a reliable prognostic indicator of favourable progression. In this study, we investigated SHM in 37 Brazilian CLL patients and identified the preferential involvement of specific immunoglobulin gene families and segments through PCR-amplified fragments or subcloned fragments. Forty-one rearrangements were observed and 37 of them were functional. A 98 percent homology cut-off with germinal sequences showed 18 patients (48.7 percent) with SHM. Unmutated cases showed a poorer clinical outcome. V H3 was the most frequent V H family, followed by V H4. The V H4-39 segment was the most frequently used, mainly in unmutated cases, while the V H3 family was predominant in mutated cases. The D3 and J H4/J H6 families were the most frequently observed.
ABSTRACT
Presentamos un paciente varón de 71 años con lesión tumoral en piel y antecedente de LLC. Los estudios histopatológicos y de inmunohistoquímica confirman el diagnóstico de Leucemia Cutis. Se realiza tratamiento con Clorambucilo y corticoides vía oral, remitiendo su enfermedad hematológica y cutánea. Actualmente luego de ocho meses del diagnóstico de LC, el paciente se encuentra libre de enfermedad.
We present a 71 year old male patient with previous records of Chronic Lymphocytic Leukaemia who presented with a tumoral skin lesion. Histological and immunohistochemical studies confirmed the Leukaemia Cutis diagnosis. The patient underwent treatment with clorambucile and systemic steroids with remision of both haemathological and skin manifestation. The patient is still under close clinical follow up and remission continues eight months so far.