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Updated: Oct 29, 2026
Isolation and Quantification of Epstein-Barr Virus from the P3HR1 Cell Line
Published on: September 28, 2022
Correlation of Epstein-Barr virus (EBV) expressed miRNAs (BART4, BART7, and BHRF1) with chronic lymphocytic leukemia
Zahra Keyvanlou1, Mehrdad Ravanshad2, Saeid Amel Jamehdar3
1Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
Background:
Epstein-Barr virus (EBV) has been shown to play a role in the pathogenesis of several B-cell malignancies, but its role in chronic lymphocytic leukemia (CLL) remains unclear. EBV-encoded microRNAs, including BHRF1 and BART families, are thought to modulate host cell survival and apoptosis. This study aimed to evaluate the expression of EBV miRNAs in CLL patients and assess their association with disease stage and viral load.
Methods:
In this study, peripheral blood mononuclear cells (PBMCs) were separated using the Ficoll-Paque density gradient method from 20 EBV-positive CLL patients, including early-stage (stage 0-1, n = 10) and advanced-stage (stage 3-4, n = 10) groups. EBV viral load was quantified using real-time PCR targeting EBNA-1, and expression levels of BHRF1, BART4, and BART7 miRNAs were measured using stem-loop RT-qPCR. Correlations between miRNA expression, viral load, and clinical stage were analyzed.
Results:
EBV viral load was significantly higher in advanced-stage CLL patients than in early-stage patients (P = 0.0363). However, miR-BHRF1 expression tended to be higher in early-stage patients than in advanced-stage patients; however, this difference was not statistically significant (P > 0.05). In contrast, miR-BART4 and miR-BART7 were significantly upregulated in advanced-stage patients (P < 0.05). Notably, BHRF1 expression positively correlated with EBV DNA load (r = 0.516, p = 0.019), while no significant association was observed for BART4 (p = 0.174, r = 0.316) or BART7 (p = 0.117, r = 0.361).
Conclusion:
Our results indicate that EBV-encoded miRNAs exhibit differential expression in CLL depending on disease stage and may contribute to leukemic progression. BHRF1, in particular, may serve as a biomarker of viral activity and early-stage disease, while BART miRNAs may promote advanced-stage pathogenesis. Further studies are warranted to validate these miRNAs as potential prognostic markers and therapeutic targets in CLL.
