Identification of the association between HBcAg-specific T cell and viral control in chronic HBV infection using a cultured ELISPOT assay

Chen, Chengcong; Jiang, Xiaotao; Liu, Xuan; Guo, Ling; Wang, Weibin; Gu, Shuqin; Wen, Chunhua; Yi, Xuan; Tang, Libo; Li, Yongyin

doi:10.1002/jlb.5ma0620-023rr

Cited by 11 publications

(15 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The straightforward comparison between sAg loss and sAg-positive patients may make it easier to identify the pivotal epitopes that may play critical roles in achieving a functional cure while neglected under various alternative epitopes in other studies. Of note, the dominant epitopes on the C protein in patients who achieved a functional cure were also consistent with our previous finding that HBV C-specific T-cell responses played an essential role in HBV control (33). To our knowledge, few studies have documented sAg loss-related dominant epitopes, which would have potentially far-reaching ramifications for immunotherapy of chronic HBV infection.…”

Section: Discussionsupporting

confidence: 90%

Identification and Mapping of HBsAg Loss-Related B-Cell Linear Epitopes in Chronic HBV Patients by Peptide Array

Liu

et al. 2021

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

Identification of immunogenic targets against hepatitis B virus (HBV)-encoded proteins will provide crucial advances in developing potential antibody therapies. In this study, 63 treatment-naïve patients with chronic HBV infection and 46 patients who achieved hepatitis B surface antigen loss (sAg loss) following antiviral treatment were recruited. Moreover, six patients who transitioned from the hepatitis B e antigen-positive chronic infection phase (eAg+CInf) to the hepatitis phase (eAg+CHep) were enrolled from real-life clinical practice. Additionally, telbivudine-treated eAg+CHep patients and relapsers or responders from an off-treatment cohort were longitudinally studied. The frequencies and function of B cells were assessed by flow cytometry. We devised a peptide array composed of 15-mer overlapping peptides of HBV-encoded surface (S), core (C), and polymerase (P) proteins and performed a screening on B-cell linear epitopes with sera. Naïve B cells and plasmablasts were increased, whereas total memory, activated memory (AM), and atypical memory (AtM) B cells were reduced in sAg- patients compared with sAg+ patients. Importantly, longitudinal observations found that AtM B cells were associated with successful treatment withdrawal. Interestingly, we identified six S-specific dominant epitopes (S33, S34, S45, S76, S78, and S89) and one C-specific dominant epitope (C37) that reacted with the majority of sera from sAg- patients. Of note, more B-cell linear epitopes were detected in CHep patients with alanine aminotransferase (ALT) flares than in nonflare CInf patients, and five B-cell linear epitopes (S4, S5, S10, S11, and S68) were overwhelmingly recognized by ALT flare patients. The recognition rates of epitopes on C and P proteins were significantly increased in CHep patients relative to CInf patients. Strikingly, a statistically significant elevation in the number of positive epitopes was observed when ALT nonflare patients shifted into the flare phase. Moreover, S76 identified at baseline was confirmed to be associated with a complete response after 48 weeks of telbivudine therapy. Taken together, we identified several functional cure-related B-cell linear epitopes of chronic HBV infection, and these epitopes may serve as vaccine candidates to elicit neutralizing antibodies to treat HBV infection.

show abstract

Section: Discussionsupporting

confidence: 90%

Identification and Mapping of HBsAg Loss-Related B-Cell Linear Epitopes in Chronic HBV Patients by Peptide Array

Liu

et al. 2021

Front. Immunol.

Self Cite

View full text Add to dashboard Cite

show abstract

“…The "ns" represents P > 0.05. Control: peptides only; polyphenol: peptides + polyphenol; iACAT: peptides + iACAT; MTA: peptides + MTA ◂ targeted epitope and stage of infection [21][22][23]. Using an adapted peptide-based expansion assay [18][19][20][21] (Fig.…”

Section: Hbv-specific T-cell Responses In Chronic Hepatitis B Infectionmentioning

confidence: 99%

“…However, such studies are limited by the low frequencies and high exhaustion status of HBV-specific T-cells in the peripheral blood mononuclear cell (PBMC) compartment. Therefore, monitoring and analysis of HBV-specific T-cells ex vivo are difficult for CHB, which usually depends on in vitro expansion [18][19][20][21] or enrichment [22][23][24]. Much effort has been devoted to developing more sensitive assays to measure HBV-specific T-cells ex vivo.…”

Section: Introductionmentioning

confidence: 99%

Metabolic interventions improve HBV envelope-specific T-cell responses in patients with chronic hepatitis B

Zhou

et al. 2023

Hepatol Int

View full text Add to dashboard Cite

Background Restoration of HBV-specific T cell immunity is a promising approach for the functional cure of chronic Hepatitis B (CHB), necessitating the development of valid assays to boost and monitor HBV-specific T cell responses in patients with CHB. Methods We analyzed hepatitis B virus (HBV) core- and envelope (env)-specific T cell responses using in vitro expanded peripheral blood mononuclear cells (PBMCs) from patients with CHB exhibiting different immunological phases, including immune tolerance (IT), immune activation (IA), inactive carrier (IC), and HBeAg-negative hepatitis (ENEG). Additionally, we evaluated the effects of metabolic interventions, including mitochondria-targeted antioxidants (MTA), polyphenolic compounds, and ACAT inhibitors (iACAT), on HBV-specific T-cell functionality. Results We found that HBV core- and env-specific T cell responses were finely coordinated and more profound in IC and ENEG than in the IT and IA stages. HBV env-specific T cells were more dysfunctional but prone to respond to metabolic interventions using MTA, iACAT, and polyphenolic compounds than HBV core-specific T-cells. The responsiveness of HBV env-specific T cells to metabolic interventions can be predicted by the eosinophil (EO) count and the coefficient of variation of red blood cell distribution width (RDW-CV). Conclusion These findings may provide valuable information for metabolically invigorating HBV-specific T-cells to treat CHB. Graphical abstract

show abstract

“…The clinical detection of host HBV-specific T cells is still very limited, since no standard T cell epitope library covering broad patients is available thus far. Most studies have utilized panels of overlapping peptides (OLPs) spanning overall HBV antigens (peptide scanning) rather than functionally validated epitope peptides (VEPs) in the ELISPOT or FluoroSpot assay for HBV-specific T cell enumeration or in the mass cytometry for T cell functional phenotype analysis, after ex vivo co-culture of OLPs pools with host lymphocytes ( 12 , 58 – 61 ). Only few researchers used a small amount of VEPs for the patient cohort with matching HLA allotype ( 62 ).…”

Section: Discussionmentioning

confidence: 99%

Screening and Identification of HBV Epitopes Restricted by Multiple Prevalent HLA-A Allotypes

Ding

Zhou

et al. 2022

Front. Immunol.

View full text Add to dashboard Cite

Although host T cell immune responses to hepatitis B virus (HBV) have been demonstrated to have important influences on the outcome of HBV infection, the development of T cell epitope-based vaccine and T cell therapy and the clinical evaluation of specific T cell function are currently hampered markedly by the lack of validated HBV T cell epitopes covering broad patients. This study aimed to screen T cell epitopes spanning overall HBsAg, HBeAg, HBx and HBpol proteins and presenting by thirteen prevalent human leukocyte antigen (HLA)-A allotypes which gather a total gene frequency of around 95% in China and Northeast Asia populations. 187 epitopes were in silico predicted. Of which, 62 epitopes were then functionally validated as real-world HBV T cell epitopes by ex vivo IFN-γ ELISPOT assay and in vitro co-cultures using peripheral blood mononuclear cells (PBMCs) from HBV infected patients. Furthermore, the HLA-A cross-restrictions of each epitope were identified by peptide competitive binding assay using transfected HMy2.CIR cell lines, and by HLA-A/peptide docking as well as molecular dynamic simulation. Finally, a peptide library containing 105 validated epitopes which cross-binding by 13 prevalent HLA-A allotypes were used in ELISPOT assay to enumerate HBV-specific T cells for 116 patients with HBV infection. The spot forming units (SFUs) was significantly correlated with serum HBsAg level as confirmed by multivariate linear regression analysis. This study functionally validated 62 T cell epitopes from HBV main proteins and elucidated their HLA-A restrictions and provided an alternative ELISPOT assay using validated epitope peptides rather than conventional overlapping peptides for the clinical evaluation of HBV-specific T cell responses.

show abstract

Identification of the association between HBcAg-specific T cell and viral control in chronic HBV infection using a cultured ELISPOT assay

Cited by 11 publications

References 44 publications

Identification and Mapping of HBsAg Loss-Related B-Cell Linear Epitopes in Chronic HBV Patients by Peptide Array

Identification and Mapping of HBsAg Loss-Related B-Cell Linear Epitopes in Chronic HBV Patients by Peptide Array

Metabolic interventions improve HBV envelope-specific T-cell responses in patients with chronic hepatitis B

Screening and Identification of HBV Epitopes Restricted by Multiple Prevalent HLA-A Allotypes

Contact Info

Product

Resources

About