Assessment of the effect of TLR7/8, TLR9 agonists and CD40 ligand on the transformation efficiency of Epstein-Barr virus in human B lymphocytes by limiting dilution assay

Younesi, Vahid; Shirazi, Forough Golsaz; Memarian, Ali; Amanzadeh, Amir; Jeddi-Tehrani, Mahmood; Shokri, Fazel

doi:10.1007/s10616-013-9542-x

Cited by 14 publications

(14 citation statements)

References 56 publications

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“…We also performed a traditional and reliable limiting dilution assay (LDA) to determine the fraction of CSCs in each subpopulation. The LDA results revealed the frequency of sphere formation, providing useful information for interpreting the composition of the mixed population of CSCs [23, 46]. Our current data showed that Lgr5-positive subpopulations possessed a higher percentage of CSCs than did Lgr5-negative subpopulations.…”

Section: Discussionmentioning

confidence: 58%

Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

Leng

Xia

Chen

et al. 2018

Cell Physiol Biochem

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Background/Aims: Although EpCAM+CD44+ cells exhibit more stem-like properties than did EpCAM-CD44- cells, the specificity of EpCAM combined with CD44 in defining CSCs needs further improvement. Lgr5 is used as a biomarker to isolate cancer stem cells (CSCs) in colorectal cancer. However, it remains unclear whether Lgr5, along with EpCAM and CD44, can further identify and define CSCs in colorectal cancer. Methods: Lgr5+CD44+EpCAM+, Lgr5+CD44+EpCAM-, Lgr5+CD44-EpCAM+, Lgr5-CD44+EpCAM+, and Lgr5-CD44-EpCAM-cells were separately isolated using fluorescence-activated cell sorting (FACS). Colony formation, self-renewal, differentiation, and tumorigenic properties of these cells were investigated through in vitro experiments and in vivo tumor xenograft models. The expression of stemness genes and CSC- and epithelial-mesenchymal transition (EMT)-related genes, such as KLF4, Oct4, Sox2, Nanog, CD133, CD44, CD166, ALDH1, Lgr5, E-cadherin, ZO-1, Vimentin, Snail, Slug, and Twist, was examined using real-time PCR. Results: Lgr5-positive subpopulations exhibited higher capacities for colony formation, self-renewal, differentiation, and tumorigenicity as well as higher expression of stemness genes and mesenchymal genes and lower expression of epithelial genes than did Lgr5-negative subpopulations. Conclusion: Our data revealed that tumorigenic cells were highly restricted to Lgr5-positive subpopulations. Most importantly, Lgr5+CD44+EpCAM+ cells exhibited more pronounced CSC-like traits than did any other subpopulation, indicating that Lgr5 combined with CD44 and EpCAM can further improve the stem-like traits of CSCs in colorectal cancer.

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Section: Discussionmentioning

confidence: 58%

Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

Leng

Xia

Chen

et al. 2018

Cell Physiol Biochem

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“…Conversely, Smad4, a well-known tumour silencer and a major regulator of intracellular TGF-β1 signalling, was up-regulated after knockdown of KLF4 expression ( Figure 1A,B). 20,24,25 Our data showed that the median frequencies were from 100/211 of CSCs-shCon cells to 100/566 of CSCs-shKLF4 cells in primary colorectal patient samples, and the median frequencies were decreased in Lgr5 + CD44 + EpCAM + cells from DLD-1 (100/484 vs 100/1304) cells after KLF4 knockdown ( Figure 1C). Because a sphere is composed of all descendants from a single CSC, the number of sphere reflects the CSC population 23 and CSC frequency can be estimated through the LDA.…”

Section: Klf4 Overexpression Facilitates Colorectal Cscs Stemness Pmentioning

confidence: 78%

Krüppel‐like factor 4 regulates stemness and mesenchymal properties of colorectal cancer stem cells through the TGF‐β1/Smad/snail pathway

Leng

Zhou

et al. 2019

J Cellular Molecular Medi

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| INTRODUC TI ONCancer metastasis contributes to approximately 90% of cancer-related deaths. 1 Accumulating evidence indicates that cancer stem cells (CSCs) are responsible for the seeding and colonization of cancer metastasis. 2-4 For the past decades, studies have explored and investigated the mechanisms of human carcinogenesis to help develop novel therapeutic strategies. To date, cancer treatment has not been much improved in advanced and chemo-resistant human cancers. Thus, a better understanding of CSCs behaviour in human cancers, including colorectal cancers (CRC), could provide insight into defining the molecular mechanisms of tumorigenesis and cancer progression. Indeed, identification of CSC markers is a very hot topic and interesting data have been generated. 5 For example, CSC markers for breast, prostate, AbstractKrüppel-like factor 4 (KLF4) was closely associated with epithelial-mesenchymal transition and stemness in colorectal cancer stem cells (CSCs)-enriched spheroid cells.Nonetheless, the underlying molecular mechanism is unclear. This study showed that KLF4 overexpression was accompanied with stemness and mesenchymal features in Lgr5 + CD44 + EpCAM + colorectal CSCs. KLF4 knockdown suppressed stemness, mesenchymal features and activation of the TGF-β1 pathway, whereas enforced KLF4 overexpression activated TGF-β1, phosphorylation of Smad 2/3 and Snail expression, and restored stemness and mesenchymal phenotypes. Furthermore, TGF-β1 pathway inhibition invalidated KLF4-facilitated stemness and mesenchymal features without affecting KLF4 expression. The data from the current study are the first to demonstrate that KLF4 maintains stemness and mesenchymal properties through the TGF-β1/Smad/Snail pathway in Lgr5 + CD44 + EpCAM + colorectal CSCs. K E Y W O R D Scancer stem cell, colorectal cancer, Krüppel-like factor 4, snail, TGF-β1

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“…Because concomitant infections can induce reactivation of the EBV lytic cycle [35], it is tempting to speculate that episodes of increased EBV load could coincide with common viral or bacterial infections inducing B cell activation in XIAP G466X patients. In a recent study, co-stimulation of cells PBMCs with Toll-like receptor (TLR)-7/8 and TLR-9 agonists increased significantly the frequency of EBV transformed B cells (P < 0·001) [36].…”

Section: Discussionmentioning

confidence: 98%

A mutation in X-linked inhibitor of apoptosis (G466X) leads to memory inflation of Epstein–Barr virus-specific T cells

López‐Granados

Stacey

Kienzler

et al. 2014

Clinical and Experimental Immunology

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SummaryMutations in the X-linked inhibitor of apoptosis (XIAP) gene have been associated with XLP-like disease, including recurrent Epstein-Barr virus (EBV)-related haemophagocytic lymphohystiocytosis (HLH), but the immunopathogenic bases of EBV-related disease in XIAP deficiency is unknown. We present the first analysis of EBV-specific T cell responses in functional XIAP deficiency. In a family of patients with a novel mutation in XIAP (G466X) leading to a late-truncated protein and varying clinical features, we identified gradual hypogammaglobulinaemia and large expansions of T cell subsets, including a prominent CD4 + CD8 + population. Extensive ex-vivo analyses showed that the expanded T cell subsets were dominated by EBV-specific cells with conserved cytotoxic, proliferative and interferon (IFN)-γ secretion capacity. The EBV load in blood fluctuated and was occasionally very high, indicating that the XIAP G466X mutation could impact upon EBV latency. XIAP deficiency may unravel a new immunopathogenic mechanism in EBV-associated disease.

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Assessment of the effect of TLR7/8, TLR9 agonists and CD40 ligand on the transformation efficiency of Epstein-Barr virus in human B lymphocytes by limiting dilution assay

Cited by 14 publications

References 56 publications

Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

Krüppel‐like factor 4 regulates stemness and mesenchymal properties of colorectal cancer stem cells through the TGF‐β1/Smad/snail pathway

A mutation in X-linked inhibitor of apoptosis (G466X) leads to memory inflation of Epstein–Barr virus-specific T cells

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