Th17 cells over 5.9% at admission indicate poor prognosis in patients with HBV-related acute-on-chronic liver failure

Abstract

Our previous study demonstrated that Th17 cells increased significantly in patients with hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF). However, their prognostic role in HBV-ACLF patients remains unknown.Sixty-eight consecutive HBV-ACLF patients were enrolled in this cohort study. Th17 cells were examined using flow cytometry. Disease severity scores were assessed. ROC curves were used to evaluate the value in predicting prognosis. Survival was analyzed using Kaplan-Meier curves. Predictors of mortality were determined by regression analysis.Th17 cells were significantly higher in HBV-ACLF patients compared to patients with chronic hepatitis B and normal controls (both P < .001). Also, Th17 cells were higher in nonsurviving HBV-ACLF patients than in surviving patients (P = .014). Th17 cells were positively correlated with CLIF-Consortium ACLF (CLIF-C ACLF) score (r = 0.240, P = .048). ROC curves showed that the frequency of Th17 cells had accuracy in predicting 90-day prognosis equivalent to MELD, MELD-Na and CLIF-C ACLF scores in HBV-ACLF (P = .34, P = .26, and P = .15, respectively). More importantly, the area under the ROC curve (AUROC) increased when Th17 cells were combined with MELD, MELD-Na or CLIF-C ACLF score than using Th17 cells alone (P = .021, P = .006, and P = .023, respectively). Kaplan-Meier analysis revealed that higher Th17 cells (≥5.9%) were closely associated with poor overall survival in HBV-ACLF (P = .0086). Additionally, multivariate regression analysis showed that the frequency of Th17 cells over 5.9% was an independent predictor of mortality (OR = 0.154, P = .025).Circulating Th17 cells positively correlated with disease severity in HBV-ACLF. The frequency of Th17 cells over 5.9% could serve as a prognostic biomarker for HBV-ACLF patients.

Figure 1

Th17 cells were stained and analyzed using flow cytometry. In this study, IL-17 + CD3 + CD8-T cells were defined as Th17 cells. Representative dotplots of IL-17 expression in peripheral CD3 + CD8- T cells of NC, CHB, and HBV-ACLF patients. The value in the upper left quadrant indicated the percentage of Th17 cells. ACLF = acute-on-chronic liver failure, CHB = chronic hepatitis B, HBV = hepatitis B virus, NC = normal control.

Figure 2

Th17 cells were significantly increased in HBV-ACLF patients independent of HBeAg presence. (A) Th17 cells were significantly higher in HBV-ACLF patients than in CHB and NC groups (both P < .001). (B) Th17 cells increased slightly in cirrhotic HBV-ACLF patients than noncirrhotic patients (P = .085). (C) No differences existed in Th17 cells between HBeAg-positive and HBeAg-negative patients in CHB group and HBV-ACLF patients. eAg-N, HBeAg-negative; eAg-P, HBeAg-positive; ^∗∗∗P < .001; ns, not significant. ACLF = acute-on-chronic liver failure, CHB = chronic hepatitis B, HBV = hepatitis B virus, NC = normal control.

Figure 3

Th17 cells were closely associated with disease severity in HBV-ACLF patients. (A) CLIF-C ACLF score is recently developed to evaluate disease severity in HBV-ACLF patients. Th17 cells were positively correlated with CLIF-C ACLF score. Also, positive correlation trends were found between Th17 cells and MELD score, between Th17 cells and MELD-Na score. (B) At admission, Th17 cells were significantly higher in patients with at least 2 complications than in patients with one complication (P < .001) or without complication (P = .025). Similarly, during the whole hospital stay, patients with at least 2 complications had higher Th17 cells at admission than those with one complication (P = .033) and without complication (P = .046). Moreover, Th17 cells were significantly higher in nonsurviving patients than in surviving patients (P = .014). Collectively, these findings indicated that Th17 cells were closely associated with disease severity in HBV-ACLF. ^∗P < .05; ^∗∗∗P < .001.

Figure 4

Higher Th17 cells at admission indicated poor prognosis in HBV-ACLF patients. The prognostic values of Th17 alone and in combination with updated prognostic parameters (MELD score, MELD-Na score and CLIF-C ACLF score) were assessed by ROC curves in HBV-ACLF patients. (A) ROC curves indicated that Th17 cells had accuracy of predicting prognosis in HBV-ACLF patients equivalent to MELD score (P = .34), MELD-Na score (P = .26) and CLIF-C ACLF score (P = .15). (B) Next, when Th17 cell levels were combined with MELD score, MELD-Na score and CLIF-C ACLF score, accuracy of predicting prognosis in HBV-ACLF patients was significantly increased than using Th17 cells alone (P = .021, P = .006 and P = .023, respectively).

Figure 5

Th17 cells over 5.9% were associated with poor overall survival in HBV-ACLF. By applying a cut-off point of 5.9%, we divided HBV-ACLF patients into the higher group (Th17 cells ≥5.9%) and the lower group (Th17 cells < 5.9%). Survival was evaluated using Kaplan–Meier curves. (A) No significant difference was found in 30-day survival between the higher and the lower groups (P = .150). (B) Importantly, 90-day survival decreased significantly in the higher group than the lower group (P = .0086).