Predictive value of Th17 and Treg cells at baseline for HBsAg loss in chronic hepatitis B patients with low HBsAg quantification treated with pegylated interferon and nucleos(t)ide analogue

Abstract

Background and aims: The primary goal of chronic hepatitis B (CHB) treatment is to reduce hepatitis B surface antigen (HBsAg). T helper 17 (Th17) and regulatory T (Treg) cells are essential for the development of CHB. However, how Th17 and Treg cells contribute to HBsAg loss is still unknown. Therefore, this study aimed to search for the predictive value of Th17 and Treg cells for HBsAg loss in CHB patients with low HBsAg quantification.

Methods: The study included 99 hepatitis B e antigen (HBeAg)-negative CHB patients who had completed a year of nucleos(t)ide analogue (NA) monotherapy and had received both NA and pegylated interferon (PEG-IFN) treatment for less than 96 weeks (96 wk). In the cured group, 48 patients lost HBsAg within 48 wk, while 51 patients did not (uncured group). Blood samples and clinical data were collected for research.

Results: During PEG-IFN and NA combination therapy, the proportion of Th17 cells in the cured group increased significantly, while the proportion of Treg cells in the uncured group increased. From 0 to 24 wk, the proportion of Th17 cells in the cured group was significantly higher than in the uncured group, while the opposite was true for Treg cells. Patients with alanine aminotransferase (ALT) ≥ 2.5 upper limit of normal (ULN) at 12 wk had a higher proportion of Th17 cells and a lower proportion of Treg cells than those with ALT <2.5 ULN at 12 wk. Additionally, the proportion of Th17 cells is inversely associated with the level of HBsAg, whereas the level of Treg cells is positively related to HBsAg quantification. The clinical cure index, including age, HBsAg quantification, and the proportions of Th17 and Treg cells, had a higher area under the curve (0.957) for predicting HBsAg loss when compared to the proportions of Th17 and Treg cells and HBsAg quantification alone.

Conclusions: Combined with quantification of HBsAg, the proportions of Th17 cells and Treg cells at baseline can be used as good predictors of HBsAg loss in patients with low HBsAg quantification treated with NA and PEG-IFN.

Keywords: Chronic hepatitis B (CHB); Clinical cure; Hepatitis B surface antigen (HBsAg) loss; Nucleos(t)ide analogue (NA); Pegylated interferon (PEG-IFN); Regulatory T (Treg) cell; T helper 17 (Th17) cell.

Fig. 1

Flow chart of the patient enrollment. Abbreviations: CHB, chronic hepatitis B; HBsAg, hepatitis B surface antigen; wk, weeks.

Fig. 2

Dynamic fluctuations of Th17 and Treg cells at baseline (0 wk), week 12 (12 wk), and week 24 (24 wk) in the cured and uncured groups and the differences of proportions of Th17 and Treg cells from 0 to 24 wk between the two groups. (A) Within 24 wk, the proportions of CD4⁺ and CD8⁺ T cells did not change significantly in the same group, and there were no significant differences between cured and uncured patients. (B) The proportions of Th17 cells (IL-17⁺CD4⁺ T cells) showed significant fluctuation within 24 wk in the cured patients. (C) In the uncured group, the proportions of Treg cells (CD25⁺CD4⁺ T cells and TGF-β⁺CD4⁺ T cells) changed significantly in 24 wk. Proportions of Th17 and Treg cells showed a substantial difference between the cured and uncured patients at (D) baseline, (E) 12 wk, and (F) 24 wk. ^∗P ＜0.05, ^∗∗P ＜0.01, ^∗∗∗P ＜ 0.001. ns, not significant. Abbreviations: IL, interleukin; TGF-β, transforming growth factor-beta; Th17, T helper 17; Treg, regulatory T.

Fig. 3

Typical graphs of flow analysis data of Th17/Treg cells for cured and uncured patients at baseline (0 wk). Abbreviations: IL, interleukin; TGF-β, transforming growth factor-beta; Th17, T helper 17; Treg, regulatory T.

Fig. 4

The differences in proportions of Th17 and Treg cells among patients with ALT ≥2.5 ULN and ALT <2.5 ULN at (A) baseline (0 wk), (B) week 12 (12 wk), and (C) week 24 (24 wk).^∗P < 0.05, ^∗∗P < 0.01. Abbreviations: ALT, alanine aminotransferase; IL, interleukin; TGF-β, transforming growth factor-beta; Th17, T helper 17; Treg, regulatory T; ULN, upper limit of normal.

Fig. 5

Proportions of IL-17⁺CD4⁺T cells and TGF-β⁺CD4⁺T cell subsets were linearly correlated with the levels of (A, B) qHBsAg and (C, D) ALT at week 12 (12 wk). Abbreviations: ALT, alanine aminotransferase; IL, interleukin; qHBsAg, quantification of hepatitis B surface antigen; TGF-β, transforming growth factor-beta.

Fig. 6

The predictive value of the proportions of Th17 and Treg cells, qHBsAg, and CCI at baseline (0 wk) for HBsAg loss. (A) The receiver operator characteristic curve of proportions of IL-17⁺/IL-21⁺/IL-10⁺/TGF-β⁺CD4⁺ T cells at baseline in predicting HBsAg loss. (B) Baseline CCI values were significantly better than qHBsAg levels in predicting clinical cure within 48 weeks. Abbreviations: AUC, area under the curve; CCI, clinical cure index; IL, interleukin; qHBsAg, quantification of hepatitis B surface antigen; TGF-β, transforming growth factor-beta; Th17, T helper 17; Treg, regulatory T.

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