Exhausted phenotype of circulating CD8+ T cell subsets in hepatitis B virus carriers - PubMed (original) (raw)

Exhausted phenotype of circulating CD8+ T cell subsets in hepatitis B virus carriers

Daixi Jiang et al. BMC Immunol. 2022.

Abstract

Background: Chronic hepatitis B virus (HBV) infection is characterized by the presence of dysfunctional exhausted CD8+ T cells that hamper viral control. We investigated the phenotypic heterogeneity of exhausted CD8+ T cells in HBV carriers.

Methods: We enrolled 31 HBV carriers and 23 healthy controls (HCs) in our study. Peripheral blood mononuclear cells (PBMCs) were isolated, and flow cytometry was used to determine the phenotypic distribution of CD8+ T cell subsets. Expression of cytokines such as TNF-α and IFN-γ was detected by quantitative reverse transcription-PCR, a fluorescence flow cytometry-based immunomicrobead assay and flow cytometry.

Results: There were no significant differences in the baseline characteristics between the 31 HBV carriers and the 23 sex- and age-matched HCs. CD8+ T cells exhibited higher levels of inhibitory receptors (TIM3 and PD1) in the HBV carriers than in the HCs (P < 0.05); in particular, Tfc cells (CXCR5+CD25-) expressed higher levels of TIM3 and PD1 than non-Tfc cells in the HBV carriers. In addition, among the subsets of Tc cells, the Tc17 (CXCR5-CD25-CCR6+) subset displayed increased expression of TIM3 and LAG3 in the HBV carriers. Our findings further showed that CD8+ T cells produced lower levels of IFN-γ, TNF-α, and Granzyme B. Paired analysis of the Tfc subset and the Tc subset indicated that higher levels of cytokines (IFN-γ and TNF-α) were produced by the Tfc subset in the HBV carriers. Among the Tc subsets, the Tc17 subset produced lower levels of cytokines.

Conclusion: The Tfc subset exhibited an enhanced exhausted phenotype but possessed some functional properties during chronic HBV infection, while the Tc subset showed a lower functional level. The identification of these unique subsets may provide a potential immunotherapeutic target in chronic HBV infection.

Keywords: CD8+ T cell; CXCR5; Exhaustion; Flow cytometry; HBV.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Fig. 1

CD8+ T cells are exhausted in HBV carriers. A Flow cytometric detection of PD1, TIM3, LAG3, and CTLA4 expression on CD8+ T cells among PBMCs from HCs and HBV carriers; B differential expression of inhibitory receptors in CD8+ T cells between HCs and HBV carriers (calculated by the percentage of IR+CXCR5+CD8+ T cells plus the percentage of IR+CXCR5−CD8+ T cells); C differential mRNA expression of TOX, PD1, TIM3, and CTLA4 in PBMCs between HCs and HBV carriers. HCs: healthy controls (n = 23), HBV: hepatitis B virus carriers (n = 31); data are presented as the means ± SEs; FDR correction was used to correct the P values for comparisons between controls and patients. *P < 0.05 and **P < 0.01

Fig. 2

CXCR5+CD8+ T cells exhibit a more suppressive phenotype than their CXCR5− counterparts in HBV carriers. A Flow cytometric detection of PD1, TIM3, LAG3, and CTLA4 on CXCR5+CD8+ T cells and CXCR5−CD8+ T cells in HBV carriers; B–E paired analysis of PD1, TIM3, LAG3, and CTLA4 expression on CXCR5+CD8+ T cells and CXCR5−CD8+ T cells in HBV carriers; F–G differential expression of PD1, TIM3, LAG3, and CTLA4 in CXCR5+CD8+ T cells and CXCR5−CD8+ T cells between HCs and HBV carriers. HCs: healthy controls (n = 23), HBV: hepatitis B virus carriers (n = 31); data are presented as the means ± SEs; *P < 0.05 and **P < 0.01

Fig. 3

Enhanced expression of inhibitory receptors on Tfc cells in HBV carriers. A Flow cytometric detection of PD1, TIM3, LAG3, and CTLA4 on Tfc and Tc/Treg cells in HBV carriers; B–E differential expression of PD1, TIM3, LAG3, and CTLA4 on Tfc, Tc, and Treg cells in HBV carriers. Tfc: follicular cytotoxic T cell (CXCR5+CD25−); Treg: regulatory T cell (CXCR5−CD25+); Tc: cytotoxic T cell (CXCR5−CD25−); HCs: healthy controls (n = 23), HBV: hepatitis B virus carriers (n = 31); data are presented as the means ± SEs; **P < 0.01; ††P < 0.01, Tfc versus Treg; ++P < 0.01, Tfc versus Tc; +P < 0.05, Tfc versus Tc

Fig. 4

Tc subsets showed heterogeneity in the suppressive phenotype in HBV carriers. A PD1 expression on Tc1, Tc2 and Tc17 cells; B TIM3 expression on Tc1, Tc2 and Tc17 cells; C CTLA4 expression on Tc1, Tc2 and Tc17 cells; D LAG3 expression on Tc1, Tc2 and Tc17 cells. HCs: healthy controls (n = 23), HBV: hepatitis B virus carriers (n = 31); data are presented as the means ± SEs; **P < 0.01

Fig. 5

The serum/mRNA levels of cytokines in PBMCs from HBV carriers. A Serum levels of cytokines as measured by the flow cytometry-based fluorescence immunomicrobead assay; B correlations between the frequencies of PD+ Tfc/Tc17 cells and the TNF-α expression level (n = 10); C correlations between the frequencies of PD+ Tfc/Tc17 cells and the IFN-γ expression level (n = 10). HCs: healthy controls, HBV: hepatitis B virus; data are presented as the means ± SEs; FDR correction was used to correct the P values for multiple testing. *P < 0.05

Fig. 6

The Tfc subset possessed higher effector functions than the Tc subset in HBV carriers. A Flow cytometric detection of IFN-γ, TNF-α, Granzyme B, and CD107a expression on CD8+ T cells among PBMCs from HCs and HBV carriers; B differential production of effector cytokines by CD8+ T cells between HCs (n = 15) and HBV carriers (n = 15); C flow cytometric detection of effector cytokines in the Tfc and Tc subsets in HBV carriers; D paired analysis of cytokine expression in the Tfc and Tc subsets in HBV carriers. HCs: healthy controls, HBV: hepatitis B virus, Tfc: follicular cytotoxic T cells (CXCR5+FOXP3−), Tc: cytotoxic T cells (CXCR5−FOXP3−); data are presented as the means ± SEs; **P < 0.01

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Exhausted phenotype of circulating CD8+ T cell subsets in hepatitis B virus carriers - PubMed (original) (raw)