{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T13:03:45Z","timestamp":1785416625533,"version":"3.56.0"},"reference-count":35,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2026,7,12]],"date-time":"2026-07-12T00:00:00Z","timestamp":1783814400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81860595"],"award-info":[{"award-number":["81860595"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["81860595"],"award-info":[{"award-number":["81860595"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100011785","name":"Natural Science Foundation of Guangxi Province, China","doi-asserted-by":"publisher","award":["2025GXNSFAA069044"],"award-info":[{"award-number":["2025GXNSFAA069044"]}],"id":[{"id":"10.13039\/501100011785","id-type":"DOI","asserted-by":"publisher"}]},{"name":"High-Level Talent Research Innovation Team Construction Project of Guangxi Center for Disease Control and Prevention, China"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Occult hepatitis B virus (HBV) infection (OBI) has the same transmission routes and clinical outcomes as overt HBV infection. However, due to the lack of reliable biomarkers, its occurrence and potential clinical harm are often overlooked. We previously found that the prevalence of OBI increases with age among individuals vaccinated at birth, but the transcriptome characteristics of their immune cells remain unknown. In this study, fifteen subjects born between 1987 and 1993 and vaccinated perinatally were recruited. They were divided into OBI, inactive chronic HBsAg carriers (ICHC) and healthy control groups. Blood samples were drawn from each subject for scRNA-seq and subsequent validation studies. The results showed that eleven cell types and nine T-cell subsets were identified, with no significant differences in the proportions of T-cell subsets among the three groups. HIPK2 expression in T cells was downregulated 1.42-fold in the OBI group versus healthy controls (p = 2.01 \u00d7 10\u221269) and upregulated 1.34-fold versus the ICHC infection group (p = 9.95 \u00d7 10\u221261). Differentially expressed genes (DEGs) in immune-related pathways were enriched in the OBI group. TP53 plays a central role in HIPK2 function in the context of OBI. Serum HIPK2 levels were significantly lower in the OBI group than in the healthy controls (p = 0.047), but they were also significantly lower than in the ICHC infection group. In conclusion, downregulated HIPK2 expression in T cells is associated with OBI. The functional role of HIPK2 in OBI, including its role as a biomarker to distinguish OBI from a healthy status, warrants further investigation.<\/jats:p>","DOI":"10.3390\/microorganisms14071523","type":"journal-article","created":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T07:28:27Z","timestamp":1783927707000},"page":"1523","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Downregulated HIPK2 Expression in T Cells Is Associated with Occult Hepatitis B Virus Infection"],"prefix":"10.3390","volume":"14","author":[{"given":"Zhi-Hua","family":"Jiang","sequence":"first","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei-Lin","family":"Huang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"},{"name":"School of Preclinical Medicine, Guangxi Medical University, Nanning 530021, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li-Ping","family":"Hu","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue-Yan","family":"Wang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Qin-Yan","family":"Chen","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lu-Juan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Zhang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ying","family":"Huang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6034-3297","authenticated-orcid":false,"given":"Tim J.","family":"Harrison","sequence":"additional","affiliation":[{"name":"Division of Medicine, University College London Medical School, London WC1E 6BT, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ge","family":"Zhong","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8628-4213","authenticated-orcid":false,"given":"Zhong-Liao","family":"Fang","sequence":"additional","affiliation":[{"name":"Guangxi Key Laboratory for the Prevention and Control of Viral Hepatitis, Guangxi Zhuang Autonomous Region Center for Disease Prevention and Control, Nanning 530028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"860","DOI":"10.4254\/wjh.v6.i12.860","article-title":"Occult hepatitis B virus infection","volume":"6","author":"Kwak","year":"2014","journal-title":"World J. Hepatol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"S64","DOI":"10.1097\/INF.0000000000002326","article-title":"Will Infant Hepatitis B Immunization Protect Adults?","volume":"38","author":"Scheifele","year":"2019","journal-title":"Pediatr. Infect. Dis. J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1855","DOI":"10.1056\/NEJM199706263362602","article-title":"Universal hepatitis B vaccination in Taiwan and the incidence of hepatocellular carcinoma in children. Taiwan Childhood Hepatoma Study Group","volume":"336","author":"Chang","year":"1997","journal-title":"N. Engl. J. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"801","DOI":"10.1002\/hep.24442","article-title":"Elimination of hepatocellular carcinoma and acute hepatitis B in children after universal infant vaccination in Alaska","volume":"54","author":"McMahon","year":"2011","journal-title":"Hepatology"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Boccalini, S., Bonito, B., Zanella, B., Liedl, D., Bonanni, P., and Bechini, A. (2022). The First 30 Years of the Universal Hepatitis-B Vaccination-Program in Italy: A Health Strategy with a Relevant and Favorable Economic-Profile. Int. J. Environ. Res. Public Health, 19.","DOI":"10.3390\/ijerph192316365"},{"key":"ref_6","unstructured":"World Health Organization (WHO) (2026, April 30). Hepatitis B. Available online: https:\/\/www.who.int\/news-room\/fact-sheets\/detail\/hepatitis-b."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"725","DOI":"10.1046\/j.1537-2995.1979.19680104098.x","article-title":"Studies of donors who transmit posttransfusion hepatitis","volume":"19","author":"Tabor","year":"1979","journal-title":"Transfusion"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Xia, R., Peng, J., He, J., Jiang, P., Yuan, C., Liu, X., and Yao, Y. (2022). The Serious Challenge of Occult Hepatitis B Virus Infection-Related Hepatocellular Carcinoma in China. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.840825"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.jinf.2019.01.002","article-title":"Vertically acquired occult hepatitis B virus infection may become overt after several years","volume":"78","author":"Eilard","year":"2019","journal-title":"J. Infect."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"952","DOI":"10.1016\/j.jhep.2020.05.042","article-title":"Occult hepatitis B Infection and hepatocellular carcinoma: Epidemiology, virology, hepatocarcinogenesis and clinical significance","volume":"73","author":"Mak","year":"2020","journal-title":"J. Hepatol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e29823","DOI":"10.1002\/jmv.29823","article-title":"Incidences of new hepatitis B infection and anti-hepatitis B core-negative occult hepatitis B infection among Japanese blood donors in relation to anti-hepatitis B surface antigen levels","volume":"96","author":"Satake","year":"2024","journal-title":"J. Med. Virol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"2351","DOI":"10.1007\/s15010-024-02290-1","article-title":"Occult hepatitis B infection in children born to HBeAg-positive women confers a low long-term risk for HBsAg-positive infection","volume":"52","author":"Eilard","year":"2024","journal-title":"Infection"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Ostankova, Y.V., Shchemelev, A.N., Serikova, E.N., Kusevitskaya, M.B., Sannikov, M.V., Gorskaya, O.A., Basina, V.V., Shirshova, N.Y., Mashkov, I.A., and Zueva, E.B. (2025). Prevalence and Molecular Characterization of Chronic and Occult Hepatitis B Virus Infection Among Pregnant Women in St. Petersburg, Russia. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26189079"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3835","DOI":"10.1016\/j.vaccine.2016.05.040","article-title":"High prevalence of active and occult hepatitis B virus infections in healthcare workers from two provinces of South Africa","volume":"34","author":"Sondlane","year":"2016","journal-title":"Vaccine"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"317","DOI":"10.1111\/jvh.13660","article-title":"Global prevalence of occult hepatitis B: A systematic review and meta-analysis","volume":"29","author":"Ji","year":"2022","journal-title":"J. Viral Hepat."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/j.jhep.2008.09.017","article-title":"Occult hepatitis B virus infection in hepatitis B vaccinated children in Taiwan","volume":"50","author":"Mu","year":"2009","journal-title":"J. Hepatol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1007\/s12072-021-10156-z","article-title":"Neonatal hepatitis B vaccination protects mature adults from occult virus infection","volume":"15","author":"Wang","year":"2021","journal-title":"Hepatol. Int."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"601","DOI":"10.1016\/j.ijid.2021.01.045","article-title":"Overt and occult hepatitis B infection after neonatal vaccination: Mother-to-infant transmission and HBV vaccine effectiveness","volume":"104","author":"Hu","year":"2021","journal-title":"Int. J. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Zhong, G., Jiang, Z.-H., Wang, X.-Y., Chen, Q.-Y., Zhang, L.-J., Hu, L.-P., Huang, M.-L., Huang, Y.-B., Hu, X., and Zhang, W.-W. (2025). Increasing Prevalence of Occult HBV Infection in Adults Vaccinated Against Hepatitis B at Birth. Vaccines, 13.","DOI":"10.3390\/vaccines13020174"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Liu, C., Wu, T., Fan, F., and Liu, Y. (2019). A portable and cost-effective microfluidic system for massively parallel single-cell transcriptome profiling. bioRxiv, bioRxiv:818450.","DOI":"10.1101\/818450"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1038\/s41587-023-01767-y","article-title":"Dictionary learning for integrative, multimodal and scalable single-cell analysis","volume":"42","author":"Hao","year":"2024","journal-title":"Nat. Biotechnol."},{"key":"ref_22","unstructured":"Gayoso, A., Shor, J., Carr, A.J., Sharma, R., and Pe\u2019er, D. (2026, April 30). GitHub: DoubletDetection. Available online: https:\/\/zenodo.org\/records\/2678042."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1007\/s00281-012-0327-7","article-title":"Occult HBV infection","volume":"35","author":"Raimondo","year":"2013","journal-title":"Semin. Immunopathol."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Jiang, X., Tian, J., Song, L., Meng, J., Yang, Z., Qiao, W., and Zou, J. (2024). Multi-omic molecular characterization and diagnostic biomarkers for occult hepatitis B infection and HBsAg-positive hepatitis B infection. Front. Endocrinol., 15.","DOI":"10.3389\/fendo.2024.1409079"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1470","DOI":"10.1053\/j.gastro.2008.02.017","article-title":"The characteristics of the cell-mediated immune response identify different profiles of occult hepatitis B virus infection","volume":"134","author":"Zerbini","year":"2008","journal-title":"Gastroenterology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"139","DOI":"10.4161\/cc.6.2.3788","article-title":"HIPK2: A versatile switchboard regulating the transcription machinery and cell death","volume":"6","author":"Calzado","year":"2007","journal-title":"Cell Cycle"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e2117112119","DOI":"10.1073\/pnas.2117112119","article-title":"HIPK2 directs cell type-specific regulation of STAT3 transcriptional activity in Th17 cell differentiation","volume":"119","author":"Cheung","year":"2022","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"eaau4604","DOI":"10.1126\/scisignal.aau4604","article-title":"HIPK2 is necessary for type I interferon-mediated antiviral immunity","volume":"12","author":"Cao","year":"2019","journal-title":"Sci. Signal."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Wang, Y., Debatin, K.M., and Hug, H. (2001). HIPK2 overexpression leads to stabilization of p53 protein and increased p53 transcriptional activity by decreasing Mdm2 protein levels. BMC Mol. Biol., 2.","DOI":"10.1186\/1471-2199-2-8"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Lim, H.Y., Han, J., Yoon, H., and Jang, K.L. (2022). Tumor Suppressor p53 Inhibits Hepatitis B Virus Replication by Downregulating HBx via E6AP-Mediated Proteasomal Degradation in Human Hepatocellular Carcinoma Cell Lines. Viruses, 14.","DOI":"10.3390\/v14102313"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1002\/1096-9071(200010)62:2<151::AID-JMV5>3.0.CO;2-N","article-title":"Occult hepatitis B virus infection in HBs antigen-negative hepatocellular carcinoma in a Japanese population: Involvement of HBx and p53","volume":"62","author":"Shiota","year":"2000","journal-title":"J. Med. Virol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Saitta, C., Pollicino, T., and Raimondo, G. (2022). Occult Hepatitis B Virus Infection: An Update. Viruses, 14.","DOI":"10.3390\/v14071504"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e37097","DOI":"10.1016\/j.heliyon.2024.e37097","article-title":"An overview of occult hepatitis B infection (OBI) with emphasis on HBV vaccination","volume":"10","author":"Delghandi","year":"2024","journal-title":"Heliyon"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1038\/cdd.2008.154","article-title":"How cells switch HIPK2 on and off","volume":"16","author":"Sombroek","year":"2009","journal-title":"Cell Death Differ."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"RP98622","DOI":"10.7554\/eLife.98622.3","article-title":"PIM kinase control of CD8 T cell protein synthesis and cell trafficking","volume":"13","author":"Marchingo","year":"2025","journal-title":"eLife"}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/7\/1523\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,13]],"date-time":"2026-07-13T07:39:32Z","timestamp":1783928372000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/14\/7\/1523"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,12]]},"references-count":35,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["microorganisms14071523"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms14071523","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,12]]}}}