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Background <p>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.</p> Methods <p>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, <i>n</i> = 10) and advanced-stage (stage 3–4, <i>n</i> = 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.</p> Results <p>EBV viral load was significantly higher in advanced-stage CLL patients than in early-stage patients (<i>P</i> = 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 (<i>P</i> &gt; 0.05). In contrast, miR-BART4 and miR-BART7 were significantly upregulated in advanced-stage patients (<i>P</i> &lt; 0.05). Notably, BHRF1 expression positively correlated with EBV DNA load (<i>r</i> = 0.516, <i>p</i> = 0.019), while no significant association was observed for BART4 (<i>p</i> = 0.174, <i>r</i> = 0.316) or BART7 (<i>p</i> = 0.117, <i>r</i> = 0.361).</p> Conclusion <p>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.</p>

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Correlation of Epstein–Barr virus (EBV) expressed miRNAs (BART4, BART7, and BHRF1) with chronic lymphocytic leukemia stages

  • 期刊论文

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  • 发表日期：

    2026年7月31日

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  • Zahra Keyvanlou,
  • Mehrdad Ravanshad,
  • Saeid Amel Jamehdar,
  • Elham Babaei

摘要

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.