[IF 21.1] Decoding the Immune Switches in Hepatitis B: How Absin Reagents Powered a Single-Cell + Spatial Multi-Omics Breakthrough

I. Research Rationale: Multi-dimensional dissection of the core logic governing immune-phenotype switching in HBV infection

HBV infection exhibits extreme clinical heterogeneity; immune status differs markedly among the immune-tolerant (IT), immune-active (IA), inactive-carrier (ICI) and acute-hepatitis-B (AHB) phases, yet the core regulatory circuitry remains incompletely defined. Adopting a “clinical-stratification → multi-omics integration → functional validation” pipeline, the authors established a comprehensive research framework:

Key-technology flow-chart:

II. Major Findings: three key discoveries in HBV immune regulation

1. Divergence of CD8⁺ T-cell subsets dictates immune-state switching

Two intra-hepatic CD8⁺ T-cell subsets were identified:

• GZMK⁺ CD8⁺ T cells: enriched in AHB and IA, exhibit liver-residency and exhaustion signatures (high CXCR6, PDCD1), mediating cytotoxicity via FasL/Fas or IFN-γ.
• GZMB⁺ CD8⁺ T cells: enriched in IT, exert cytotoxicity through perforin and inversely correlate with clinical inflammatory indices.

Pseudotime trajectory analysis further revealed three HBV-specific CTL (hpCTL) states: FASLG⁺ State-3 dominates IA, whereas GZMB⁺PRF1⁺ State-1 dominates IT, offering stage-specific therapeutic targets (Fig. S3a, S8).

2. A multi-cellular interaction network governs immune tolerance versus activation

• Hepatocytes: peri-portal Hep-2 subset in IT shows higher HBV infection frequency, high MHC-I but low co-stimulatory molecules (CD40, CD80), driving CD8⁺ T-cell dysfunction.
• Macrophages: MRC1⁺CDH5⁺ Kupffer cells up-regulate antigen-presentation (HLA, TAP1) and co-stimulatory molecules, enhancing IL-2 signalling in AHB/IA and rescuing CD8⁺ T-cell function.
• Regulatory circuits: PGE2-EP4 signalling is activated in IA (plasma PGE2 positively correlates with ALT), suppressing CD8⁺ T-cell cytotoxicity; IL-33⁺ liver-sinusoidal-endothelial cells and DC-SIGN⁺ macrophage-mediated type-2 immunity are markedly enhanced in IA (Fig. S5, S6).

3. Clinical relevance: immune subsets correlate with disease progression

Exhausted CD8⁺ T cells, Tregs and CXCL13⁺ Tfh cells positively correlate with plasma ALT and histological inflammation grades, whereas GZMB⁺ CD8⁺ T cells show inverse correlation, offering potential biomarkers for HBV prognosis (Fig. S11).

III. Absin Products Empower Key Experiments: core tools for multiplex IHC

In this study multiplex fluorescent IHC was pivotal for mapping intra-hepatic cell localisation and phenotypes, and was reliably supported by two Absin reagents:

1. Product list

Product	Cat. No.	Application
TSA 5-color kit	abs50013	Multiplex fluorescent IHC staining
Citrate-EDTA Antigen Retrieval Solution	abs9249	Antigen retrieval in FFPE sections

2. Role in the study

3. Key panels supported by Absin reagents

Panel	Content	Absin contribution
Fig. S2c	Fluorescent staining of CD8⁺ T-cell subsets (GZMB⁺, GZMK⁺) in liver	abs9249 exposes epitopes; abs50013-20T enables simultaneous five-colour detection
Fig. S4i	Spatial localisation of AREG, EGFR, etc.	TSA amplification secures detection of low-abundance targets
Fig. S1b/d	Clustering and phase-specific proportions of intra-hepatic cells	mIHC validates scRNA-seq annotation

IV. Absin – Your reliable partner in scientific discovery

This multi-dimensional dissection of HBV immune regulation would not have been possible without precise and robust experimental tools. Absin’s TSA multiplex kits and antigen-retrieval reagents, recognised for consistent performance and high efficiency, empowered the team to achieve accurate multi-marker detection in liver tissue, providing critical technical support for the published conclusions.

From immune-phenotyping and tumour-microenvironment profiling to infectious-disease immunology, Absin remains committed to supplying premium research tools. We continuously optimise our IHC, flow-cytometry and molecular-biology portfolios to help scientists overcome experimental bottlenecks and accelerate translational research.

Moving forward, Absin will keep pace with cutting-edge technologies and launch innovative products tailored to emerging needs, standing side-by-side with the global research community to explore the mysteries of life and advance medicine.

Products used in this paper

Cat. No.	Name	Size
abs50013	5-color multiplex fluorescent IHC staining kit	20T/50T/100T
abs9249	Citrate-EDTA antigen retrieval solution	100 mL

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