Dendritic cells (DCs), as professional antigen-presenting cells, play a pivotal role as critical mediators bridging innate and adaptive immunity. Human DCs are broadly categorized into three main types, including monocyte-derived DCs (moDCs), plasmacytoid DCs (pDCs), and conventional DCs (cDCs) (6). MoDCs, characterized by the surface markers CD14, FcγRI (CD64), and FcεRI, become activated primarily under inflammatory conditions (7). In contrast, pDCs are identified by their expression of CD123, CD303 (BDCA2), and CD304 (BDCA4). These cells specialize in robust type I interferon (IFN-I) production in response to single-stranded viral RNA and DNA, a function mediated through pattern recognition receptors (PRRs) such as Toll-like receptor (TLR)-9 (8). The cDC population, often referred to as myeloid DCs (mDCs) in literature, consists of two principal subsets: cDC1s and cDC2s. cDC1s express CD141 (BDCA3) and excel at cross-presenting exogenous antigens on MHC class I molecules to activate CD8⁺ T cells. Conversely, cDC2s, which express CD1c/BDCA1 and CD172a, primarily present antigens on MHC class II molecules to stimulate CD4⁺ T cells (7).

Emerging evidence reveals profound DC dysfunction during chronic HBV infection, with distinct pathophysiological manifestations across disease phases. Studies demonstrate reduced mDC frequencies alongside elevated B7-H1 (PD-L1) expression on mDCs in chronic hepatitis B (CHB) patients (9, 10). Similarly, decreased peripheral pDC percentages, reduced TLR9 expression, and impaired CpG-induced IFN-α responses are observed in CHB patients compared to healthy controls (HCs) (11–15). Notably, Ouaguia et al. have reported higher pDC frequencies in CHB livers than those in HCs, while liver cDCs remain comparable (16). This dysfunction extends to disrupted crosstalk between pDCs and natural killer (NK) cell, evidenced by impaired cytotoxic activation of NK cells in CHB patients (17). Beyond classical DC subsets, recent studies by Li et al. have identified expanded circulating follicular DCs (FDCs; CD14⁺ CD21^high)in chronic HBV patients compared to HCs (18).

Both circulating and intrahepatic cDC2s from HBV-infected patients exhibit reduced CD40/CD80 expression, whereas peripheral and hepatic pDCs display elevated CD40 levels compared to HCs (16). Altered expression of co-stimulatory/co-inhibitory molecules on DCs is prominent in CHB that co-stimulatory molecules (OX40L and 4-1BBL) are downregulated on peripheral pDCs and cDC1s, while PD-L1 expression on cDC2s and pDCs inversely correlates with HBV DNA (16). CD86 expression on pDCs is elevated in both immune-tolerant (IT) and immune-active (IA) phases compared to controls, with IA patients showing higher CD86 levels and enhanced IFN-α2 production (19). In addition, TGF-β1 significantly elevate within intrahepatic cDC2s and pDCs of IT patients compared to other disease stages or HCs (20). Metabolic disturbances are also evident, as Dumolard et al. have recently demonstrated dysregulated glycolysis and oxidative phosphorylation (OXPHOS) in hepatic cDC1s and pDCs across HBV infection stages (20). Furthermore, peripheral DCs from IT patients show significantly reduced levels of free cholesterol, lipid rafts, and LDL receptor (LDLR) compared to HCs. This lipid raft impairment, potentially influenced by HBsAg, can be partially restored by lipophilic statins, which also enhances the antigen-presentation ability of DCs. (21). Improtantly, functional recovery of DCs emerges in disease resolution phases, with inactive carriers (IC) demonstrating superior DC functionality over IT patients through increased expression of CD80, CD86, HLA-DR and IL-12 (22).

Functional impairments are further highlighted by TLR stimulation assays. Chronic HBV patients show significantly reduced production of IL-12p40/70 and TNFα by cDC2s, IFNα/TNFα/IFNλ1 by pDCs, and IFNλ1/TNFα/IL-12p40/70 by cDC1s compared to HCs (16). In contrast, intrahepatic DCs from CHB patients retain full functionality upon TLR triggering, producing pro-inflammatory cytokines at levels comparable to HCs (16). Furthermore, study on purified peripheral moDCs from CHB patients reveals heightened activation. The expression of both MHCII and co-stimulated molecules (CD80, CD86) as well as the cytokines (TNF-α, IL-10, IL-12) secretion in the purified peripheral moDCs from CHB patients are significantly higher than those from HCs when co-cultured with supernatant of HepG2.2.15 cells (23). Interestingly, enhanced autophagy is also observed in mo-DCs from chronic HBV patients compared to healthy donors upon re-exposure to HBV (23).

The immunomodulatory effects of antiviral therapies on DC populations exhibit substantial heterogeneity across clinical studies ( Table 1 ). One study has illustrated that entecavir (ETV) therapy significantly reduces B7-H1 expression on peripheral DCs in CHB patients through suppression of HBcAg-mediated AKT/ERK/p38 signaling pathways (9). Furthermore, another study has shown that ETV induces early pDC proliferation (12-24 weeks), while CD86 is downregulated on pDCs in HBV DNA non-responders (27). Six-month therapy of adefovir dipivoxil (ADF) restores mDC frequency and enhances their capacity to produce TNF and IL-12, whereas the frequency and TNF-α and IL-10 secretion of pDCs remain refractory (26).

Interferon-based regimens reveal distinct immunostimulatory patterns. PEG-IFN-α-2a treatment induces sustained CD86 upregulation on pDCs in patients achieving functional cure (24). Moreover, the frequency of pDC increases at week 24 post-treatment in the functional cure group (24). Consistently, Cao et al. have found that HBsAg decline significantly associates with CD86 elevation on pDCs during IFN-α treatment (27). Mechanistically, IFN-α treatment enhances hepatic pDC expansion and upregulates TLR-9 mRNA in peripheral blood mononuclear cells (PBMCs) of virological responders (25). A recent clinical trial of selgantolimod (TLR8 agonist) has demonstrated significant increase of peripheral pDCs, with a dose-dependent trend (29). Taken together, critical analysis identifies three determinants of DC functional restoration, including baseline DC subset characteristic, different antiviral agents, and variant duration of treatment and observation.

Monocytes, originating from common myeloid progenitors (CMPs) in the bone marrow, constitute approximately 10% of human peripheral leukocytes and perform multifaceted functions in homeostasis and inflammation (30). In humans, two functionally distinct subsets are recognized. CD14⁺⁺ CD16^- “migratory” monocytes are capable of tissue infiltration, and CD14⁺ CD16⁺ “patrolling” monocytes maintain vascular surveillance (31).

Chronic HBV exposure induces immunoregulatory reprogramming of monocytes. Monocytes from chronically infected individuals demonstrate elevated expression of TNF-α, IL-10, TGF-β, PD-L1, Gal-9 and HLA-E compared to HCs (32, 33). Notably, PD-L1 upregulation on monocytes is particularly pronounced in HBeAg-positive patients (34, 35). Furthermore, the hepatic compartment of chronic HBV (CHB) patients shows an enrichment of monocytes expressing Gal-9 and PD-L1 compared to HCs (33). Functionally, monocytes from IT patients and HBeAg-positive or -negative CHB patients demonstrate suppressed signaling through TLR2, TLR4, and TLR9 compared to ICs and HCs. This functional impairment is accompanied by reduced production of IL-12, TNF-α, and IL-6, as well as diminished phagocytic capacity and oxidative response (36–38). Moreover, PD-L1- and Gal-9-expressing monocytes in CHB contribute to the dysregulation of both adaptive and innate immune responses (33). Another study has revealed significantly downregulated expression of membrane-bound CD163, a monocyte activation marker, on circulating monocytes from both treatment-naïve CHB patients and those achieving HBsAg loss compared to HCs (37). Conversely, circulating soluble CD163 (sCD163) levels are elevated in CHB patients with significant inflammation (A≥2) or fibrosis (F≥2) (37).

Emerging evidence suggests antiviral interventions may partially reverse HBV-induced monocyte dysfunction, though therapeutic outcomes remain heterogeneous. After one year of treatment, tenofovir disoproxil fumarate (TDF) fails to restore monocyte functionality, as evidenced by unchanged monocyte subset distribution and proportions expressing PD-LI, Gal-9, TLR-2, IL-12, IL-10, CD64, and iNOS before and after treatment (33, 38), whereas responders to Peg-IFN-α and ETV demonstrate partial TLR9 expression recovery on monocytes (36). Intrahepatic transcriptomics reveal elevation of hepatic monocytes after 24-week PEG-IFN-α treatment (39). Recent single-cell analyses reveal that PEG-IFN-α reduces proportions of pro-inflammotory CD14⁺ and CD16⁺ monocytes, accompanied by systemic immune reprogramming from TNF-α-dominant to IFN-α-driven transcriptional profiles (40). Consistently, NUC-treated patients exhibit upregulated expression of TLR-associated genes LY6E and STK4 on monocytes compared to ICs (41). A recent clinical trial of selgantolimod (TLR8 agonist) has demonstrated significant increase of peripheral CD14⁺ classical monocytes, with a dose-independent trend (29). Collectively, these findings position monocytes as pivotal mediators of HBV immunopathogenesis. While current antivirals show partial efficacy in reversing monocyte dysfunction, stratified interventions targeting subset-specific reprogramming are needed to achieve functional cure.

Myeloid-derived suppressor cells (MDSCs), constituting less than 1% of myeloid cells in healthy individuals (42), are a heterogeneous population of immunosuppressive myeloid cells comprising two functionally distinct subsets, polymorphonuclear MDSCs (PMN-MDSCs) and monocytic MDSCs (M-MDSCs) (43, 44). In human PBMCs, these subsets are phenotypically characterized as CD11b⁺ CD14⁻ CD15⁺/CD66b⁺ (PMN-MDSC) and CD11b⁺CD14⁺HLA-DR^−/loCD33⁺CD15⁻ (M-MDSC) (45). MDSCs undergo significant expansion under pathological conditions, suppressing T cell responses and promoting disease progression through multiple mechanisms (46).

Several clinical studies demonstrate remarkable expansion of circulating MDSCs in CHB patients compared to HCs (47–49) The frequency of MDSCs positively correlates with HBV DNA load, HBeAg levels and HBsAg levels (48, 50). Both M-MDSC and granulocytic-MDSC (gMDSCs) from different phases of CHB expressed high TGF-β and IL-10 (51). Notably, purified M-MDSCs from HBeAg-positive patients exhibit enhanced suppression of CD4⁺/CD8⁺ T cell proliferation and IFN-γ production compared to those from HBeAg-negative individuals (52). Moreover, gMDSCs expressing arginase expand during high viral replication phases, impairing T cell function via arginase-dependent pathways (53). Notably, an enrichment of PD-L1/Arg/iNOS expressing hepatic MDSCs is observed in CHB patients compared to HCs (51). A recent single-cell RNA sequencing of PBMC has shown that a CD14⁺ cluster with an MDSC-like phenotype predominantly accumulates in patients with CHB, with high expression of genes with immunoregulatory functions (54).

Apart from peripheral immune suppression, MDSCs also contribute to central tolerance via chemokine-mediated trafficking. HBsAg upregulates CCR9 expression on M-MDSCs through ERK1/2-IL-6 signaling, facilitating thymic homing via CCL25 chemotaxis (55). This process enables peripheral HBsAg transport to thymic medulla, ultimately inducing clonal deletion of HBsAg-specific CD8⁺ thymocytes, a mechanism predominant in pediatric CHB patients (55). Collectively, these findings unveal MDSCs as central orchestrators of HBV-induced immune tolerance through peripheral and thymic mechanisms, offering potential targets for therapeutic intervention.

Current evidence suggests suboptimal efficacy of NUCs in reconstituting MDSC homeostasis. One-year TDF monotherapy fails to restore MDSC frequency and the secretion of IL-10 and TGF-β or improve HBV-specific T-cell responses (51, 56). Strikingly, patients achieving functional cure through PEG-IFN-α-2a display substantial M-MDSC reduction (57). Consistently, targeting MDSCs with all-trans retinoic acid restores HBV-specific CD4⁺ and CD8⁺ T cell proliferation and IFN-γ production in CHB patients (50).

As critical effectors of innate immunity, NK cells mediate rapid antiviral and antitumor responses. In humans, NK cell populations are traditionally classified into CD56^dim (cytotoxic) and CD56^bright (immunoregulatory) subsets based on CD56 and CD16 surface marker expression (58). NK cells exhibit dual roles in HBV immunity, balancing antiviral defense mechanisms and immunopathogenic potential through liver injury (59). During acute HBV infection (AHB), peripheral CD56^bright NK cells undergo significant expansion (60) and display an activated phenotype characterized by upregulated activation receptors (NKp30, NKp44, NKp46 and NKG2C), activation markers (CD38 and HLA-DR), and cytotoxic mediators like TRAIL, alongside downregulation of inhibitory receptors (CD158a/b and NKG2A) (61). Elevated CD107a expression and robust IFN-γ production upon IL-12+ IL-18 or K562 stimulation have also been observed in peripheral CD56^bright NK cells during acute HBV (61). Notably, CD56^dim NK cell-mediated antibody-dependent cellular cytotoxicity (ADCC) associates with early HBsAg clearance in AHB (62). Temporal analyses, however, reveal transient suppression of IFN-γ and TNF-α production during peak viremia, with functional recovery upon viral resolution (63).

In chronic HBV infection, phenotypic and functional defects of NK cells are well-documented. Discrepancies in circulating NK cell frequencies across studies reflect population heterogeneity and clinical phase variations (64–66). Progressive NK cell dysfunction has been observed in chronic HBV infection, characterized by reduced expression of activating receptors (e.g. NKG2D), increased inhibitory checkpoint molecules (PD-1, Tim-3, CD94) (67), with the frequency of intrahepatic PD-1⁺ NK cells being the highest in HBeAg+ HBV patients (68). This dysfunction is further marked by attenuated antiviral cytokine (IFN-γ, TNF-α) secretion (69, 70), and elevated immunosuppressive IL-10/TGF-β1 production (71, 72). Conversely, NK cells may negatively regulate HBV-specific T cells through TRAIL-R2-mediated lysis (73). Furthermore, the activation of NK cells driven by proinflammatory cytokines (IFN-α, IL-12, IL-15, IL-8) also exacerbates liver inflammation via NKG2D/TRAIL/IFN-γ-mediated hepatocyte damage, particularly in IA phase (64, 74–76). This pathogenic role is supported by a positive correlation between intrahepatic NK cell accumulation and histological inflammation severity (77). Furthermore, TRAIL expression on CD56^bright NK cells positively correlates with liver inflammation and ALT flare (65, 71, 75). Intrahepatic analyses of a recent single-cell RNA sequencing demonstrate that the CXCR6+ NCAM1+ CD160^high liver-resident NK-cell cluster with a significant higher expression of IL-32 within the HBsAg-high group compared to HBsAg-low group (78).

Functional analyses reveal discrepancies in NK cell cytotoxic activity. While NK cells from IA patients exhibit enhanced TNF-α, IFN-γ, and CD107a production compared to HCs (75, 79), cytokine-mediated functional exhaustion has been reported following IL-2 and IL-12 or IL-21 stimulation (71, 80, 81). Conversely, other studies illustrate preserved cytotoxic function of NK cells, as evidenced by intact K562 lysis capacity (82, 83).

Emerging evidence reveals the multifaceted immunomodulatory effects of antiviral therapies on NK cells in CHB ( Table 2 ). NUCs and PEG-IFN-α therapies have demonstrated marked heterogeneity across studies regarding capacity to reshape NK cell quantity, phenotype, and function, influenced by treatment duration, therapeutic agents, and patient-specific factors. A recent randomized controlled trial has observed significant upregulation of activation markers (TRAIL, HLA-DR, Ki-67, CD38) and receptors (NKp46, NKG2D, NKp30, NKG2A) on total NK cells—irrespective of HBsAg decline magnitude (98). However, some studies report transient expansion of immunoregulatory CD56^bright subset during NUC therapy, with normalization post-HBsAg clearance (88, 96), while other investigations document static or even reduced NK cell counts in NUC-treated cohorts, including telbivudine (LDT) and ETV (81, 97). Intrahepatic transcriptomics reveal no alteration of hepatic NK cells after 24-week PEG-IFN-α treatment (39).

The phenotype of NK cells varies among different studies following antiviral therapy. Inhibitory receptors such as NKG2A and KIR2DL3 demonstrate progressive downregulation in tandem with viral suppression under NUC therapy (88, 90). Consistently, activation receptors(NKp30, NKp46, and NKG2D) exhibit temporal upregulation patterns that parallel HBsAg clearance trajectories (88, 92, 94, 95). ETV monotherapy transiently suppresses NKG2D and NKp30 expression on NK cells in HBeAg-positive patients (85), whereas therapeutic regimen switching (ADV to ETV) enhances CD244⁺ activated NK subsets (86). PEG-IFN-α induces TRAIL upregulation on CD56^bright NK cells in complete responders (102), while LAM-ADV combination therapy restores TRAIL expression without rescuing IFN-γ production deficits in CD56^dim subsets (71). Intriguingly, ETV treatment enhances CD69 expression and IFN-γ production specifically within CD56^bright NK populations (81). Furthermore, PEG-IFN-α discontinuation in plateau-phase patients reduces exhaustion markers (CD57, TIGIT) on CD56^dim NK (93). Several clinical trials of novel immunotherapies exhibit prominent alteration on NK cells. GS-9620 (TLR-7 agonist) rapidly upregulates NK activation markers (CD69, TRAIL, HLA-DR) and enhances effector functions (IFN-γ, TNF-α, degranulation) (99). Preliminary research of selgantolimod (TLR-8 agonist) activates NK cells as well, evidenced by CD69 expression (100), while another phase II trial demonstrates no alteration in circulating NK cell frequencies (29). α-GalCer modulates NK cell frequencies bidirectionally (decreasing at lower doses, increasing at 10 μg/kg), and effectively increases CD69 expression (101). Collectively, these findings highlight the critical role of NK cells in antiviral immunity, with treatment-induced phenotypic remodeling potentially serving as a biomarker for therapeutic efficacy.

MAIT cells are characterized by their semi-invariant TCR α-chain (usually Vα7.2–Jα33/12/20 in humans) and restriction to the MHC-I-related protein MR1 (103, 109), which presents microbial riboflavin (vitamin B2) and folate (vitamin B9) derivatives (110). MAIT cells constitute approximately 5% of circulating T cells (111) but are enriched in mucosal tissues, representing up to 45% of hepatic T lymphocytes (109). Upon activation, they predominantly secrete IFN-γ and TNF, with a minor subset producing IL-17A (109).

The frequency, phenotype and cytokine production of MAIT cells exhibits conflicting patterns across studies in CHB patients. Several studies have reported reduced circulating MAIT cells in CHB compared to HCs (112, 113), whereas another study documents comparable levels (114). MAIT cell reduction is also observed in patients with HBV-related acute-on-chronic liver failure (115). Mechanistically, this reduction potentially attributes to conjugated bilirubin-mediated apoptosis of MAIT cells (113). Several studies have documented the upregulation of activation markers (CD69, HLA-DR, CD38), immunosenescence marker CD57, and inhibitory receptors (PD-1, CTLA-4) on peripheral MAIT cells in CHB compared to HCs (113, 116, 117). However, another study demonstrates reduced expression of CD69 on MAIT cells in CHB patients (118). Notably, CD69 expression on MAIT cells correlates positively with HBV viral load, while inhibitory markers (PD-1 and CTLA-4) on MAIT cells show negative correlation with HBV DNA levels (115, 116). Further functional assessments show enhanced IFN-γ and Granzyme B secretion from MAIT cells in CHB patients than those in HCs upon anti-CD28/E.coli co-stimulation (114, 118), whereas combined stimulation of IL-12 and IL-18 yields impaired IFN-γ responses in CHB patients (119). Single-cell transcriptomics identify two hepatic MAIT subsets in CHB, T7(CD3⁺SLC4A10⁺TNFAIP3⁺) cells displaying proinflammatory cytokine secretion and immune cell recruitment capacities, and T6(CD3⁺SLC4A10⁺TNFAIP3^-) cells with impaired antiviral function (120). The progressive shift toward T6 predominance during advanced hepatic inflammation highlights MAIT cell dysfunction in chronic HBV pathogenesis (120). These findings collectively illustrate the complex duality of MAIT cell responses in CHB, balancing protective immunity with inflammation-driven exhaustion. Longitudinal analyses suggest preserved MAIT cell frequencies during NUC therapy (114, 121). Nevertheless, treatment-induced normalization of CD38 activation marker expression implies partial recovery of MAIT cell functionality, though complete phenotypic and functional restoration remains to be established (114). A phase 1b clinical trial of selgantolimod (TLR8 agonist) shows the elevation of CD69 on MAIT after a single dose (100).

γδ T cells are defined by their unique TCR consisting of a γ-chain and a δ-chain, which enables antigen recognition independent of MHC class I/II molecules (103). Two major subsets exist in humans, Vδ1⁺ and Vδ2⁺ γδ T cells (122). Vδ1+ cells, characterized by pairing of the Vδ1 chain with diverse Vγ family members (Vγ2/3/4/5/8/9) (123) predominantly reside in mucosal and epithelial tissues such as intestinal epithelium (124), skin (125, 126), spleen and liver. In contrast, Vδ2⁺ cells typically express an invariant Vγ9 chain paired with Vδ2 (127), constituting 50–95% of circulating γδ T cells in human peripheral blood (128, 129). These cells are activated through phosphoantigen recognition via butyrophilin 3A1 (BTN3A1) (130, 131), triggering rapid secretion of cytotoxic molecules and Th1 cytokines (IFN-γ and TNF-α) to combat malignancies and microbial pathogens (132, 133). Additionally, Vδ3⁺ T cells have been found in the periphery which only consist about 0.2% of γδ T cells, while in the liver they are more abundant. Limited studies on this subset show their capacity to secret Th1, Th2 and Th17 cytokines (134).

Acute HBV infection significantly reduces peripheral γδ T cell proportions and absolute counts compared to CHB and HCs, negatively correlating with serum ALT (135). AHB patients exhibit heightened activation profiles in circulating γδ T cells compared to HCs, characterized by upregulated CD38, HLA-DR, granzyme B, CD107a, and distinct transcriptional polarization as Tbet^+/hi Eomes^dim Vδ1 subsets and Tbet^dim Eomes^hi Vδ2 subsets (135, 136). Concurrently, intrahepatic γδ T cells accumulate in inflamed liver lobules during AHB (135), a phenomenon recapitulated in acute HBV murine models where hepatic γδ T cell expansion coincides with early-stage IFN-β production (137).

In chronic HBV infection, peripheral γδ T cells are significantly reduced in CHB patients relative to HCs (138), particularly in severe liver inflammation (ALT>3×ULN) (139). However, one study reports comparable γδ T cell frequencies between symptomatic CHB and HCs (140), and some studies document elevated Vδ1 T cell percentages in CHB (138, 141). Hepatic γδ T cells, particularly the Vδ2 subset, decrease in CHB patients, especially within the IA group (138). Analysis of paired samples further reveals markedly lower hepatic Vδ2 T cell levels than their peripheral counterparts in IA patients (138).

The phenotype and function of γδ T cells varies among different studies. Elevated exhaustion markers (PD-1, Tim-3 and Lag-3) and activation markers (CD69, CD38 and HLA-DR) levels are frequently reported in CHB (140, 142). Paradoxically, Chang et al. have observed decreased PD-1, CD38, Ki-67, Tim-3, and CD158a expression on Vδ2 T cells from CHB patients compared to HCs (136). Intriguingly, PD-1 expression on circulating Vδ2⁺ cells inversely correlates with serum 25(OH)D3 levels in CHB (142). PMA/ionomycin stimulation enhances IFN-γ/granzyme B/TNF-α co-expression on γδ T cells from CHB patients (136, 141). However, another study describes suppressed IFN-γ secretion of γδ T cells, but can be reversible by Tim-3/Lag-3 blockade (136, 140). Functional cytotoxicity assays reveal impaired γδ T cell-mediated lysis of HBV-infected hepatocytes in symptomatic CHB compared to HCs, though asymptomatic carriers retain partial cytolytic activity than symptomatic patients (139).

The impact of antiviral therapies on γδ T cell populations in CHB treatment presents complex immunological modifications ( Table 3 ). A randomized controlled trial has revealed TDF/PEG-IFN-α combination therapy in HBV-suppression patients exhibits no significant alterations in γδ T cell frequencies or their functional capacity to produce IFN-γ/TNF-α/granzyme B/CD107a (143). Conversely, PEG-IFN-α monotherapy reduces γδ T cell numbers, accompanied by enhanced TNF-α/CD107a expression (144, 145). Furthermore, treatment responders exhibit distinct γδ T cell differentiation patterns characterized by transient early effector cell expansion and reduced T_em subsets (145). Longitudinal monitoring of LDT therapy suggests that elevated baseline CD4⁻CD8⁻ γδ T cells predict non-response and virologic relapse (146). Collectively, these findings underscore the heterogeneity of γδ T cell responses during CHB therapy, with dynamic changes in subsets and functional markers correlating with treatment efficacy and relapse risk.

Natural killer T (NKT) cells constitute a specialized lymphocyte population distinguished by their recognition of lipid antigens presented through the CD1d molecule (103). These CD1d-restricted cells are broadly classified into two subsets, invariant NKT (iNKT) cells and diverse (type II) NKT cells. iNKT cells are characterized by a semi-invariant TCR architecture, featuring a conserved α chain rearrangement Vα24-Jα18 paired with limited β chain diversity Vβ11 in humans (103). This unique TCR configuration enables iNKT cells to detect both endogenous and exogenous lipid antigens, including the prototypical α-galactosylceramide (α-GalCer), presented via the MHC-I-like CD1d molecule (103). Additionally, iNKT cells can be activated in a TCR-independent manner through innate cytokines like IL-12 and IL-18 (147). In contrast to their invariant counterparts, type II NKT cells possess highly diverse αβ TCR repertoires while maintaining CD1d-restricted lipid antigen specificity (148). Current understanding of type II NKT cell functionality remains limited, though emerging evidence suggests their involvement in both immunoregulatory and pathogenic responses through distinct lipid antigen recognition pathways (149).

Chronic HBV infection markedly alters homeostasis and function of iNKT cells. Both peripheral and hepatic iNKT cells are significantly reduced in CHB patients compared to HCs, with negative correlation between circulating iNKT cell counts and liver injury severity (143, 150, 151). Furthermore, CD4^- iNKT cells are reduced in CHB, especially in those with detectable HBV DNA levels (151). Functional analyses reveal complex dysregulation of CD1d-iNKT axis in chronic HBV infection. Despite hepatic CD1d upregulation, the CD1d-iNKT system remains unactivated in CHB, showing impaired α-Galcer responses (150, 152). Surface marker profiling unveils a complex phenotype characterized by increased expression of NKG2A (153) and activation markers (CD69, CD38, HLA-DR) (150) alongside elevated exhaustion markers (Tim-3, PD-1) and reduced CD28 co-stimulation in both peripheral and hepatic iNKT cells from CHB patients compared to HCs (154). However, one study reports no significant upregulation in circulating or hepatic iNKT populations (150). Functional restoration is achieved in vitro through Tim-3/PD-1 blockade or CD28 activation (154). Other studies reveal that enhanced chemokine receptor expression (CCR5 and CCR6) and elevated Fas and FasL levels on peripheral iNKT cells from CHB (150). Moreover, IFN-γ⁺ NKT cells positively correlated with ALT levels and inversely correlated with HBV DNA (79, 155). Besides, other studies report diminished IL-4 and IFN-γ production in iNKT cells from CHB patients, partially reversible by exogenous IL-2 and IL-15 (150, 153, 154), while other studies find comparable cytokine production post-stimulation across disease phases upon stimulation of α-GalCer and PMA (156, 157).

Antiviral therapies elicit heterogeneous modulation of NKT cells ( Table 4 ). PEG-IFN-α add-on TDF therapy reduces peripheral iNKT frequencies without altering cytokine profiles (IFN-γ and TNF-α) (143). However, other studies observe that PEG-IFN-α or LDT monotherapy conversely increase iNKT frequencies (156, 158). Notably, baseline iNKT frequencies predict sustained response to PEG-IFN-α monotherapy in HBeAg-positive patients (158). ETV treatment differentially modulates iNKT subsets, enhancing IFN-γ⁺ while reducing IL-4⁺ iNKT cells during six-month treatment (155). LDT therapy selectively reduces peripheral CD3⁺CD56⁺ NKT-like cells in treatment responders instead of non-responders (97). Longitudinal analyses further reveal post-treatment expansion of circulating CD4⁻ iNKT subsets, with baseline elevations in the CD4⁻/CD4⁺ iNKT cell ratio correlating with HBeAg seroconversion (156). Novel immunotherapies reveal distinct mechanisms. Oral HBV envelope proteins trigger a >2-fold increase in iNKT frequency alongside improved histology and seroconversion (159), while α-GalCer administration transiently suppresses total NKT cells at 2 days post-injection (recovering by day 7) and drives a shift toward CD8⁺ predominance, most prominently at the 1 μg/kg dose (101). Collectively, these findings highlight the heterogeneity of NKT cell responses across therapeutic regimens, emphasizing NKT cells as potential biomarkers for therapeutic stratification and outcome prediction in CHB management.