Mitochondria-related parameters of lymphocyte subsets can distinguish different disease stages in patients with HBV infection

Abstract

To explore the difference of peripheral immune indexes especially T subsets cell mitochondrial indexes in patients with chronic HBV infection (CHB), liver cirrhosis (LC), hepatocellular carcinoma (HCC) and healthy controls (HCs). From May 2023 to December 2023, HBV infected patients aged 40-59 years (HBV infection group, 236 cases) and healthy people of the same age group (control group, 112 cases) were selected as the study objects. More than 80 parameters related to lymphocyte including cell percentage, absolute count, mitochondrial mass and mitochondrial low membrane potential were obtained by flow cytometry. The mitochondrial mass of memory T cells was found to be sequentially raised in the disease progression of hepatitis B patients from CHB to LC and eventually to HCC, and mitochondrial low membrane potential ratio was sequentially decreased. Then we screened out 4 main indicators related to the stage of disease progression in HBV-infected patients and distinguished the differences among the control group, CHB, LC, and HCC groups through the calculation of MPOLS (mitochondrial parameters of lymphocyte subsets) value, which can be used as a novel biomarker for hepatitis B disease progression. Mitochondrial parameters of peripheral lymphocytes in HBV-infected patients are significant indicators of different stages of disease progression. The MPOLS provides rich clinical data support for clinical early warning of high-risk hepatitis B patients, and we believe that the MPOLS can be combined with a variety of clinical characteristics to achieve more accurate diagnosis and treatment.

Keywords: HBV infection; Lymphocyte subsets; Mitochondrial low membrane potential; Mitochondrial mass.

Fig. 1

The distribution characteristics of samples and the significant difference of pair-to-pair comparison. (A) The sample distribution map shows the age/gender distribution of the samples. (B) Venn diagram of different mitochondrial parameters between CHB, LC, HCC and HCs. Red circles show 33 strongly correlated indicators. (C) An UpSet diagram showing pairwise comparison of differentially mitochondrial parameters between CHB, LC, HCC, and HCs. The figure shows the number of common cells mitochondrial parameters in different combinations. The lower left corner shows groups with differential parameters and their number that were screened among each group. The dots on the right show parameters common or specific to the corresponding group. The histogram shows the number of specific or common parameters. Most of all, black points mean that the parameter between the two groups have significant differences, while gray points mean nonsignificant. The red area shows 33 strongly correlated indicators. Venn diagram and UpSet diagram were completed by BioLadder software (https://www.bioladder.cn/web/#/pro/cloud).

Fig. 2

The mitochondrial parameters of memory T cells were different in CHB, LC, HCC patients and HCs group. The Venn chart shows the difference between patients at CHB, LC and HCC stages of hepatitis B with the HCs control group (A), and the difference between CHB, LC and HCC groups (B). Heat maps showing mitochondrial parameters (Mitochondrial mass, MM; Mitochondrial low membrane potential ratio, MMP^low%) showed differences between the control group and the HBV disease group (C). The mean fluorescence intensity (MFI) stands for the mitochondrial mass of memory T cells in HCs, CHB, LC and HCC group (D–H lower), and the significant difference of MMP^low% in the HCs, CHB, LC and HCC group (D-H upper).

Fig. 3

The main parameters of MPOLS were obtained by random forest analysis and their differences among groups were analyzed. (A) Four main parameters (T4Tem.MMP^low%; T8Tem.PD-1⁺.Abs.Count; (CD3⁺)T. MMP^low%; CD3⁻CD56⁺Abs.Count) of MPOLS were obtained by random forest analysis. (B) The violin/box plot of MPOLS value in the HCs, CHB, LC and HCC group. (C) Receiver operating characteristics (ROC) curves describe the performance of the prediction of MPOLS for different clinical outcomes in HBV-infected patients. Numbers in the color key represent the area under the ROC curve (AUC).

Fig. 4

Chronic hepatitis B virus infection and MPOLS in relation to liver disease progression. The figure shows the clinical stages involved in the natural history of CHB, and the risk of complications in this disease progression. MPOLS value provides valuable predictive data for the effective management of CHB. It is important to monitor patients at high risk and to treat them early to prevent liver complications, cirrhosis, and HCC development.