Galectin-9 and myeloid-derived suppressor cell as prognostic indicators for chronic lymphocytic leukemia.

BACKGROUND: Galectin-9 and myeloid-derived suppressor cells (MDSCs) have an important role in tumors, but their clinical values in chronic lymphocytic leukemia (CLL) have not been fully elucidated. This study aimed to analyze the prognosis values of Galectin-9 and MDSCs in CLL. METHODS: The concentrations of Galectin-9, argininase-1, and inducible nitric oxide synthase in serum were detected by enzyme-linked immune sorbent assay. The expression of Tim-3 protein in peripheral blood mononuclear cell was detected by Western blot. Flow cytometry was used to analyze the percentages of Tim-3 on T-cells (CD3+ T, CD4+ T, and CD8+ T cells) and MDSCs. RESULTS: Our results showed that Galectin-9 and MDSCs significantly increased in CLL patients and were closely related to the disease progression. Patient's receiver operating characteristic, progression-free survival, and Cox regression analysis showed that Galectin9 and MDSCs were poor prognostic factors of CLL. CONCLUSION: Galectin-9 and MDSCs were associated with clinical progression and could be important prognostic indicators for CLL.

Expression and Clinical Significance of Helper T Cells 9 and Its Sytokines Interleukin 9 in Chronic Lymphocytic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the expression and clinical significance of T helper cell 9 (Th9) and its cytokine interleukin 9(IL-9) in peripheral blood of patients with chronic lymphocytic leukemia(CLL).@*METHODS@#A total of 43 newly diagnosed patients with chronic lymphocytic leukemia in the First Affiliated Hospital of Xinjiang Medical University from June 2021 to June 2022 were selected as the case group. The patients were divided into Binet A group (13 cases), Binet B group (20 cases) and Binet C group (10 cases) by Binet staging system, and 20 healthy volunteers who underwent physical examinationin in our hospital in the same period served as control group. The proportion of Th9 cells in peripheral blood was detected by flow cytometry, the expression level of Th9 specific transcription factors PU.1 and IRF4 was detected by Western blot, and the expression level of serum cytokine IL-9 was detected by ELISA. The proportion of Th9, the expression of PU.1, IRF4 and IL-9 in each group were compared, and the correlation between the proportion of Th9, IL-9 and clinicopathological indexes of CLL patients was analyzed.@*RESULTS@#The proportion of Th9, the expression of PU.1, IRF4 and IL-9 in CLL group were significantly higher than those in control group (P<0.05), the proportion of Th9 and the expression of IL-9 in Binet B and C group were higher than those in Binet A group (P<0.05), but there was no significant difference in the proportion of Th9 cells between Binet B group and C group (P>0.05). The expression of IL-9 in Binet C group was significantly higher than that in Binet B group (P<0.05) . The proportion of Th9 cells and IL-9 were highly expression in patients with β2 microglobulin abnormality, IGHV unmutation, P53 abnormality and hepatosplenic lymph node enlargement(P<0.05), but not related to age and sex (P>0.05). The results of Spearman correlation analysis showed that the proportion of Th9 in patients with CLL was negatively correlated with the lymphocytic account and lymphocyte proportion(rs=-0.32，rs=-0.34). The proportion of Th9 and IL-9 were positively correlated with Binet stage, Rai stage and CLL-IPI Scoring (rs=0.79，rs=0.54，rs=0.58； rs=0.72，rs=0.63，rs=0.45), but not with WBC, CD4+ T cells and CD8+T cells (P>0.05). The proportion of Th9 was positively correlated with IL-9 (rs=0.53).@*CONCLUSION@#Th9 cells and IL-9 are abnormally highly expressed in CLL, which is related to the poor prognosis of CLL.

Activated Galectin-9/Tim3 promotes Treg and suppresses Th1 effector function in chronic lymphocytic leukemia.

Tim-3 is a negative immunoregulator in anti-tumor response, but its mechanism in chronic lymphocytic leukemia (CLL) is not yet clear. The aim of this study was to understand the role of Galectin-9/Tim-3 signaling pathway in the regulation of CD4+ T cell subsets in CLL patients. Flow cytometry results showed that the number of Treg cells obviously increased, and there was a significant Treg/Th17 imbalance in CLL patients. In addition, Tim-3 overexpressed on the surface of Th1 and Treg cells in CLL patients. The levels of Galectin-9 and IL-10 were significantly elevated in patients of CLL, especially in stages of Binet B, and C. However, IFN-γ decreased. Moreover, Galectin-9 in CLL patients was positively correlated with the number of Tim-3+ Treg cells and the level of IL-10. Interestingly, when the Tim-3/Galectin-9 pathway was blocked in vitro, the level of IL-10 in the culture supernatant of CD4+ T was significantly reduced, while the levels of IFN-γ and TNF-α were increased. After co-culture with activated Th1 cells, the apoptosis of CLL cells was significantly increased, and this effect was reversed after treatment with Tim-3+ Tregs. In summary, Galectin-9/Tim-3 are elevated in CLL and associated with disease progression. By the negative regulation of CD4+ T cells, activated Galectin-9/Tim-3 suppresses Th1 effector function and also promotes Treg to be involved in immune escape of CLL. This pathway might become the potential target of immunotherapy in CLL patients.

Expressions of T-cell immunoglobulin mucin-3 and galectin-9 in acute lymphoblastic leukemia and their clinical significances / 白血病·淋巴瘤

Objective:To explore the expression levels, clinical significances and prognostic evaluation value of T-cell immunoglobulin mucin-3 (Tim-3) and galectin-9 (Gal-9) in bone marrow cells of patients with newly diagnosed acute lymphocytic leukemia (ALL).Methods:Bone marrow samples from 30 newly diagnosed ALL patients admitted to First Affiliated Hospital of Xinjiang Medical University from September 2016 to September 2018 were selected, and peripheral blood samples from 20 healthy volunteers during the same period in the First Affiliated Hospital of Xinjiang Medical University were treated as the controls. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect mRNA relative expression levels of Tim-3 and Gal-9. Differences in mRNA expression of Tim-3 and Gal-9 among ALL patients with varied clinicopathological characteristics were compared. Overall survival (OS) analysis was performed by using the Kaplan-Meier method, Cox proportional hazards model was used to make univariate and multivariate survival analysis.Results:mRNA relative expression levels of Tim-3 and Gal-9 in 30 newly diagnosed ALL patients were higher than those in the healthy control group (2.86±0.47 vs. 0.45±0.05, t = 21.65, P<0.05; 9.79±0.58 vs. 0.96±0.23, t = 63.24, P<0.05). mRNA relative expression level of Tim-3 had statistically significant differences in patients with different ages, France-America-Britain (FAB) Cooperative Group classification, hazard grades and central nervous system invasion (all P<0.01). There were statistically significant differences in mRNA relative expression level of Gal-9 for patients with different ages, FAB Cooperative Group classification, white blood cell count (WBC), central nervous system invasion and NOTCH1 mutation (all P<0.01). All patients were grouped by mRNA relative expression levels of Tim-3 and Gal-9, and patients in high Tim-3 expression group (≥2.86) had worse overall survival (OS) compared with that for patients in low Tim-3 expression group (<2.86) ( P = 0.048). Patients in high Gal-9 expression group (≥9.79) had worse OS compared with that for patients in low Gal-9 expression group (<9.79) ( P = 0.031). Moreover, the OS in Tim-3 and Gal-9 both high expression group was worse than that in Tim-3 and Gal-9 both low expression group and in the low expression group of either of them (all P<0.05). There was no statistically significant difference in OS between the high Tim-3 expression with low Gal-9 expression group and the high Gal-9 expression with low Tim-3 expression group ( P > 0.05). Multivariate Cox regression analysis revealed that peripheral blood WBC≥11.4×10 9/L, BCR-ABL gene mutation, central nervous system invasion, and high expression of Tim-3 and Gal-9 were independent risk prognostic factors of OS for newly diagnosed ALL patients (all P<0.05) . There was a positive correlation between the expression levels of Tim-3 and Gal-9 ( r = 0.788, P<0.01). Conclusions:The high expression of Tim-3 and its ligand Gal-9 are independent effecting factors of poor prognosis in newly diagnosed ALL patients. The expression levels of Tim-3 and Gal-9 can be served as a potential prognostic indicator for ALL patients.