Immunomodulation and Molecular Diagnostic Aspects in Individuals with Hepatitis B Virus Infection
DOI:
https://doi.org/10.31530/cjnst.2026.2.2.8Keywords:
Viral Load, HBeAg, IFN-γ, TNF-α, CD8, RT-PCR, HLA-A, HBcAgAbstract
Background: Hepatitis B virus (HBV) is a major global health issue associated with severe liver complications. Outcomes depend on the immune response, which can sometimes fail, and are associated with chronic disease. Improved diagnostics and understanding of immune markers are crucial for more effective management and treatment of HBV infection.
Aims: The study aimed to investigate the association between the HBV viral load, viral antigens, including Hepatitis B surface antigen (HBsAg), Hepatitis B core antigen (HBcAg), Hepatitis B enveloped antigen (HBeAg), and pro-inflammatory cytokines, including Human Interferon γ (INF-γ), Human Tumour Necrosis Factor α (TNF-α), Soluble Human Leukocyte Antigen class I (sHLA-A), an antigen-presenting molecule, and Human soluble Cluster of Differentiation 8 (sCD8), a soluble T-cell activation marker. This study provides additional regional information on the associations between virological and immunological parameters in patients with HBV infection from the Kurdistan Region of Iraq.
Methodology: The study was a case-control study conducted on 120 individuals in the Kurdistan Region of Iraq, comprising 60 patients with HBV infection and 60 healthy controls. The investigation included both sexes aged 25 to 74 years, during the period from March to July 2025. RT-PCR was used to assess viral load, and ELISA was used to evaluate immunological markers.
Results: According to the study findings, Patients with HBV infection exhibited significantly higher immunological markers than healthy controls (p < 0.0001). Significant positive correlations were observed in the study between viral load and HBeAg (r=0.9051), as well as inflammatory cytokines IFN-γ (r=0.9189), TNF-α (r=0.832), sCD8 (r=0.888), and HLA-A (r=0.801). The obtained results demonstrated significant positive correlations between viral load and immune activation markers.
Conclusion: Overall, the current study demonstrated significant positive correlations among viral load, viral antigens, and immunological markers in patients with chronic HBV infection and provides additional evidence of the immunological characteristics.
Downloads
References
[1] A. F. Shallal, "Immunomodulatory Pathways of Some Biomarkers in Individuals with Renal Failure," Aro-the Scientific Journal of Koya University, 2025, vol. 13, no.(2), p. 326–333, DOI: https://doi.org/10.14500/aro.12474
[2] A. F. Shallal, "Modulation of immune pathway and bi-omarkers: impacts of lifestyle, nutrition, and therapeutic interventions," Journal of Pharmaceutical Research, 2025, vol. 14, no.(24), p. 419–430, DOI: https://doi.org/10.5281/ZENODO.17949376
[3] A. F. Shallal, "Association of Immunomodulation with Biological Agents: A Review," World Journal of Phar-maceutical Research, 2026, vol. 15, no.(6), p. 849–860, DOI: https://doi.org/10.5281/ZENODO.19045651
[4] A. F. SHALLAL, C. H. RAZAQ, and M. N. OWAID, "Immunomodulatory Effects of Ganoderma lucidum on NK Cells: From Traditional Use to Modern Immuno-therapy," Journal of Macrofungi, 2025, vol. 1, no.(1), p. 14–29, DOI: https://doi.org/10.65999/macrofungi.2025.27
[5] A. F. Shallal, "Immune System Modulation: The Roles of Leukocytes, Vitamin D, Plant Extracts, Nanotech-nology, and Biomarkers in Disease and Health," World Journal of Pharmaceutical Research, 2026, vol. 15, no.(3), p. 250–263, DOI: https://doi.org/10.5281/ZENODO.18427459
[6] C. Seeger and W. S. Mason, "Molecular Biology of Hepatitis B Virus Infection," Virology, 2015, vol. 479, p. 672–686, DOI: https://doi.org/10.1016/j.virol.2015.02.031
[7] J. E. Song, "Diagnosis of hepatitis B," Annals of Trans-lational Medicine, 2016, vol. 4, no.(18), p. 338, DOI: https://doi.org/10.21037/atm.2016.09.11
[8] G. C. Fanning, F. Zoulim, J. Hou, and A. Bertoletti, "Therapeutic strategies for hepatitis B virus infection: towards a cure," Nature Reviews Drug Discovery, 2019, vol. 18, no.(11), p. 827–844, DOI: https://doi.org/10.1038/s41573-019-0037-0
[9] P. J. Easterbrook, N. Luhmann, S. Bajis, M. S. Min, M. Newman, O. Lesi, and M. C. Doherty, "WHO 2024 hepatitis B guidelines: an opportunity to transform care," The lancet Gastroenterology & hepatology, 2024, vol. 9, no.(6), p. 493–495, DOI: https://doi.org/10.1016/S2468-1253(24)00089-X
[10] W. H. Organization, Global hepatitis report 2024: ac-tion for access in low-and middle-income countries. 2024: World Health Organization.
[11] S. Sabeena and N. Ravishankar, "Horizontal modes of transmission of hepatitis B virus (HBV): a systematic review and meta-analysis," Iranian Journal of Public Health, 2022, vol. 51, no.(10), p. 2181, DOI: https://doi.org/10.18502/ijph.v51i10.10977
[12] Y. Shi and M. Zheng, "Hepatitis B virus persistence and reactivation," BMJ, 2020, vol. 370, DOI: https://doi.org/10.1136/bmj.m2200
[13] P. Zheng, Y. Dou, and Q. Wang, "Immune response and treatment targets of chronic hepatitis B virus infec-tion: innate and adaptive immunity," Frontiers in Cellu-lar and Infection Microbiology, 2023, vol. 13, p. 1206720, DOI: https://doi.org/10.3389/fcimb.2023.1206720
[14] Y.-P. Liu and C.-Y. Yao, "Rapid and quantitative detec-tion of hepatitis B virus," World Journal of Gastroenter-ology, 2015, vol. 21, no.(42), p. 11954–11963, DOI: https://doi.org/10.3748/wjg.v21.i42.11954
[15] A. Bertoletti and P. T. Kennedy, "The immune tolerant phase of chronic HBV infection: new perspectives on an old concept," Cellular & Molecular Immunology, 2015, vol. 12, no.(3), p. 258–263, DOI: https://doi.org/10.1038/cmi.2014.79
[16] M. K. Maini and A. R. Burton, "Restoring, releasing or replacing adaptive immunity in chronic hepatitis B," Nature Reviews Gastroenterology & Hepatology, 2019, vol. 16, no.(11), p. 662–675, DOI: https://doi.org/10.1038/s41575-019-0196-9
[17] S. Donica, C. N. Ciurea, M. A. Budianu, A. Cighir, R. L. Coșeriu, A. M. Bunea, and A. Man, "Correlation be-tween HBV viral load and other paraclinical parameters in patients with chronic hepatitis B," Acta Marisiensis-Seria Medica, 2025, vol. 71, no.(3), p. 223–226, DOI: https://doi.org/10.2478/amma-2025-0036
[18] [18] T.-C. Tseng, C.-J. Liu, C.-Y. Hsu, C.-M. Hong, T.-H. Su, W.-T. Yang, C.-L. Chen, H.-C. Yang, Y.-T. Huang, and S. F.-T. Kuo, "High level of hepatitis B core–related antigen associated with increased risk of hepatocellular carcinoma in patients with chronic HBV infection of intermediate viral load," Gastroenterology, 2019, vol. 157, no.(6), p. 1518–1529. e3, DOI: https://doi.org/10.1053/j.gastro.2019.08.028
[19] G. L. H. Wong, "Updated guidelines for the prevention and management of chronic hepatitis B—World Health Organization 2024 compared with China 2022 HBV guidelines," Journal of Viral Hepatitis, 2024, vol. 31, p. 13–22, DOI: https://doi.org/10.1111/jvh.14032
[20] F. S. Ali, M. H. Nguyen, R. Hernaez, D. Q. Huang, J. Wilder, A. Piscoya, T. G. Simon, and Y. Falck-Ytter, "AGA clinical practice guideline on the prevention and treatment of hepatitis B virus reactivation in at-risk in-dividuals," Gastroenterology, 2025, vol. 168, no.(2), p. 267–284, DOI: https://doi.org/10.1053/j.gastro.2024.11.008
[21] N. Le Bert, A. Koffas, R. Kumar, F. Facchetti, A. T. Tan, U. S. Gill, L.-Y. Mak, S. W. Stretch, T. Miao, and S. Hariharaputran, "Rapid whole-blood immune profil-ing reveals heterogeneous HBV-specific T cell response patterns independent of clinical disease phases," Journal of Hepatology, 2025, vol. 83 no.(6), p. 1292–1304, DOI: https://doi.org/10.1016/j.jhep.2025.06.012
[22] X. Yuan, Y. Shao, R. Huang, S. Seery, H. Wang, N. Hu, L. Wen, X. Lin, and L. Zhang, "Understanding the influence of cytokines in intrauterine hepatitis B trans-mission: A cross-sectional study in China," Cytokine, 2024, vol. 181, p. 156670, DOI: https://doi.org/10.1016/j.cyto.2024.156670
[23] Y. Shu, S. Li, Y. Du, and X. Zheng, "Anti-HBV treat-ment partially restores the dysfunction of innate im-mune cells and unconventional T cells during chronic HBV infection," Frontiers in Immunology, 2025, vol. 16, p. 1611976, DOI: https://doi.org/10.3389/fimmu.2025.1611976
[24] S. H. Hosseini Khorami, F. Nejatollahi, and M. A. Da-varpanah, "Serum Levels of Interleukin-4, Interleukin-10 and Interferon-γ in Patients with Chronic Hepatitis B Infection," Hepatitis Monthly, 2018, vol. 18, no.(4), DOI: https://doi.org/10.5812/hepatmon.60377
[25] T. Thu, "Serum Level of IL-6, TNF-Α, IL-4, IL-10 in Patients with Cirrhosis Due to Chronic Hepatitis B," Vietnam Journal of Infectious Diseases, 2024, vol. Vol. 4 No. 36 (2021), DOI: https://doi.org/10.59873/vjid.v4i36.206
[26] M. Holmberg, H. C. D. Aass, O. Dalgard, E. Samuel-sen, D. Sun, N. K. Björkström, A. Johannessen, and D. H. Reikvam, "Treatment cessation in HBeAg-negative chronic hepatitis B: clinical response is associated with increase in specific proinflammatory cytokines," Scien-tific Reports, 2023, vol. 13, no.(1), p. 22590, DOI: https://doi.org/10.1038/s41598-023-50216-y
[27] H. Li, S. Xiao, C. Huo, S. Yang, J. Wang, X. Lan, M. Li, L. Shi, L. Zhuo, and J. Zhang, "Targeting ferropto-sis restores the antiviral activity of CD8+ T cells during chronic HBV infection," Cellular and Molecular Gas-troenterology and Hepatology, 2025, p. 101612, DOI: https://doi.org/10.1016/j.jcmgh.2025.101612
[28] F. Andreata, K. D. Moynihan, V. Fumagalli, P. Di Lu-cia, D. C. Pappas, K. Kawashima, I. Ni, P. H. Bessette, C. Perucchini, and E. Bono, "CD8 cis-targeted IL-2 drives potent antiviral activity against hepatitis B virus," Science Translational Medicine, 2024, vol. 16, no.(729), p. eadi1572, DOI: https://doi.org/10.1126/scitranslmed.adi1572
[29] K. Heim, Sagar, Ö. Sogukpinar, S. Llewellyn-Lacey, D. A. Price, F. Emmerich, A. R. Kraft, M. Cornberg, S. Kielbassa, and P. Knolle, "Attenuated effector T cells are linked to control of chronic HBV infection," Nature Immunology, 2024, vol. 25, no.(9), p. 1650–1662, DOI: https://doi.org/10.1038/s41590-024-01928-4
[30] A. Letafati, O. S. Ardekani, M. Naderisemiromi, M. Norouzi, M. Shafiei, S. Nik, and S.-H. Mozhgani, "Un-ravelling the dynamic mechanisms of natural killer cells in viral infections: insights and implications," Virology Journal, 2024, vol. 21, no.(1), p. 18, DOI: https://doi.org/10.1186/s12985-024-02287-0
[31] A. Tălăngescu, M. Tizu, B. Calenic, D. F. Mihăilescu, A. E. Constantinescu, and I. Constantinescu, "HLA ge-netic diversity and chronic hepatitis B virus infection: effect of heterozygosity advantage," Medical Sciences, 2024, vol. 12, no.(3), p. 44, DOI: https://www.mdpi.com/2076-3271/12/3/44#
[32] U. Maimunah, P. Wardhani, C. D. K. Wungu, A. F. Rahmanda, A. A. Kurniawan, and A. Palayukan, "Cor-relation between quantitative HBsAg and quantitative HBV DNA in chronic hepatitis B patients: a systematic review and meta-analysis," Egyptian Liver Journal, 2024, vol. 14, no.(1), p. 35, DOI: https://doi.org/10.1186/s43066-024-00336-5
[33] B. Garba, A. A. Mohamud, F. H. Mohamed, A. A. Moussa, Y. Yakubu, N. I. Dirie, A. M. Abdulatif, J. H. Mohamoud, A. S. Ali, and A. H. Jibril, "Prevalence, risk factors, and viral load quantification of HBV infec-tion among hospital patients: A cross-sectional investi-gation in Mogadishu, Somalia," PLoS One, 2026, vol. 21, no.(1), p. e0340800, DOI: https://doi.org/10.1371/journal.pone.0340800
[34] K.-N. Tsai and J.-H. J. Ou, "Hepatitis B virus e antigen and viral persistence," Current Opinion in Virology, 2021, vol. 51, p. 158–163, DOI: https://doi.org/10.1016/j.coviro.2021.10.003
[35] Y. Qiu, Q. Tang, X.-Q. Liu, Y.-L. Xue, Y. Zeng, and P. Hu, "Hepatitis B core-related antigen as a promising serological marker for monitoring hepatitis B virus cure," World Journal of Hepatology, 2025, vol. 17, no.(1), p. 98658, DOI: https://doi.org/10.4254/wjh.v17.i1.98658
[36] L. O. Downs, M. Delphin, M. van Schalkwyk, S. Hugo, S. F. Lumley, E. Waddilove, T. Wang, J. Martin, C. de Lara, and A. Babbs, "Evaluation of Hepatitis B core re-lated antigen (HBcrAg) as a biomarker in cohorts from the United Kingdom and South Africa," Journal of In-fection, 2025, vol. 91, no.(3), p. 106601, DOI: https://doi.org/10.1016/j.jinf.2025.106601
[37] Z. Zhu, J. Wu, J. Li, and T. Wu, "Challenges and Solu-tions in pgRNA Measurement: Toward Improved Moni-toring of Hepatitis B Therapy," Pathogens, 2026, vol. 15, no.(2), p. 153, DOI: https://www.mdpi.com/2076-0817/15/2/153#
[38] L. I. Bozomitu, A. Lupu, V. V. Lupu, N. Gimiga, D. T. Anton Paduraru, D. E. Mîndru, M. Mihai, C. Anton, E. Anton, and M. Mitrea, "Accuracy of Diagnostic Inves-tigations in Monitoring Hepatitis B Virus Infection: Strengths, Limitations, and Emerging Biomarkers," In-ternational Journal of Molecular Sciences, 2026, vol. 27, no.(5), p. 2464, DOI: https://www.mdpi.com/1422-0067/27/5/2464#
[39] J. Lin, S. Jiang, X. Chen, M. Zhu, and H. Zhang, "The significance of detecting HBV pgRNA and HBcrAg in HBV patients treated with NAs," Medicine, 2024, vol. 103, no.(14), p. e37752, DOI: https://doi.org/10.1097/MD.0000000000037752
[40] T. Inoue, T. Watanabe, and Y. Tanaka, "Hepatitis B core-related antigen: A novel and promising surrogate biomarker to guide anti-hepatitis B virus therapy," Clin Mol Hepatol, 2023, vol. 29, no.(4), p. 851–868, DOI: https://doi.org/10.3350/cmh.2022.0434
[41] Y. Liu, D. Wu, K. Zhang, R. Ren, Y. Liu, S. Zhang, X. Zhang, J. Cheng, L. Chen, and J. Huang, "Detection technology and clinical applications of serum viral products of hepatitis B virus infection," Frontiers in Cellular and Infection Microbiology, 2024, vol. 14, p. 1402001, DOI: https://doi.org/10.3389/fcimb.2024.1402001
[42] B. Jiang, L. Wang, H. Liu, L. Wang, R. Su, L. Xu, G. Wei, J. Li, F. Lu, and X. Chen, "Association of HBV serological markers with host antiviral immune response relevant hepatic inflammatory damage in chronic HBV infection," Journal of Medical Virology, 2024, vol. 96, no.(4), p. e29569, DOI: https://doi.org/10.1002/jmv.29569
[43] M. Manea, I. Mărunțelu, and I. Constantinescu, "Ex-tended analysis on peripheral blood cytokines correlated with hepatitis B virus viral load in chronically infected patients–a systematic review and meta-analysis," Fron-tiers in Medicine, 2024, vol. 11, p. 1429926, DOI: https://doi.org/10.3389/fmed.2024.1429926
[44] L. Xiao, K. Tang, T. Fu, X. Yuan, S. Seery, W. Zhang, Z. Ji, Z. He, Y. Yang, and W. Zhang, "Cytokine pro-files and virological markers highlight distinctive im-mune statuses, and effectivenesses and limitations of NAs across different courses of chronic HBV infec-tion," Cytokine, 2024, vol. 173, p. 156442, DOI: https://doi.org/10.1016/j.cyto.2023.156442
[45] S. Lens, A. R. Burton, J. Davies, M. Locatelli, M. Gar-cía-López, A. Pocurull, A. Jeffery-Smith, N. Novikov, S. P. Fletcher, and X. Forns, "Tracking B cell immunity during perturbation of hepatitis B infection induced by treatment withdrawal," Gut, 2025, p. 1–10, DOI: https://doi.org/10.1136/gutjnl-2024-333309
[46] Y. Wang, J. Gu, G. Chen, Y. Jiang, Y. Xu, X. Huang, W. Sun, and J. Gan, "Roles of inflammatory cytokines in the pathogenesis of hepatitis B virus-related acute-on-chronic liver failure and CAR-T therapy," Virology Journal, 2025, vol. 22, no.(1), p. 281, DOI: https://doi.org/10.1186/s12985-025-02868-7
[47] L. Begré, A. Boyd, M.-L. Plissonnier, B. Testoni, C. Béguelin, F. Suter-Riniker, C. Scholtès, J. K. Rock-stroh, K. Lacombe, and L. Peters, "Circulating HBV RNA and hepatitis B core-related antigen as determi-nants of HBsAg loss in persons with HIV in Europe," JHEP Reports, 2025, vol. 8, no.(2), p. 101671, DOI: 10.1016/j.jhepr.2025.101671
[48] U. S. Gill, V. J. Appleby, and S. Lens, "What is new in hepatitis B," Frontline Gastroenterology, 2026, vol. 17, no.(2), p. 161–171, DOI: 10.1136/flgastro-2024-102850
[49] [M. G. Ghany, M. Buti, P. Lampertico, and H. M. Lee, "Guidance on treatment endpoints and study design for clinical trials aiming to achieve cure in chronic hepatitis B and D: Report from the 2022 AASLD-EASL HBV-HDV Treatment Endpoints Conference," Hepatology, 2023, vol. 78, no.(5), p. 1654–1673, DOI: https://doi.org/10.1016/j.jhep.2023.06.002
[50] H. Ma, Q.-Z. Yan, J.-R. Ma, D.-F. Li, and J.-L. Yang, "Overview of the immunological mechanisms in hepati-tis B virus reactivation: Implications for disease pro-gression and management strategies," World Journal of Gastroenterology, 2024, vol. 30, no.(10), p. 1295, DOI: https://doi.org/10.3748/wjg.v30.i10.1295
[51] A. Schuch, A. Hoh, and R. Thimme, "The role of natu-ral killer cells and CD8+ T cells in hepatitis B virus in-fection," Frontiers in Immunology, 2014, vol. 5, p. 258, DOI: https://doi.org/10.3389/fimmu.2014.00258
[52] J. P. Costa, A. de Carvalho, A. Paiva, and O. Borges, "Insights into immune exhaustion in chronic hepatitis B: A review of checkpoint receptor expression," Pharma-ceuticals, 2024, vol. 17, no.(7), p. 964, DOI: https://www.mdpi.com/1424-8247/17/7/964#
[53] J. Lin and A. J. Gehring, "Role of Immune Cells in Hepatitis B Virus and Associated Sequelae," Annual Review of Pathology: Mechanisms of Disease, 2026, vol. 21, p. 399–421, DOI: https://doi.org/10.1146/annurev-pathmechdis-042524-030646
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Charmo Journal of Natural Sciences and Technologies
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
