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Isolation and Expansion of Cytotoxic Cytokine-induced Killer T Cells for Cancer Treatment
Published on: January 24, 2020
Profiling the dynamic expression of checkpoint molecules on cytokine-induced killer cells from non-small-cell lung
Lin Zhang1,2,3, Jian Wang1,2,3, Feng Wei1,2,3
1Department of Immunology, Tianjin Medical University Cancer Institute and Hospital, Tiyuanbei, Tianjin, China.
Abstract:
Immune checkpoints associate with dysfunctional T cells, which have a reduced ability to clear pathogens or cancer cells. T-cell checkpoint blockade may improve patient survival. However, checkpoint molecules on cytokine-induced killer (CIK) cell, a non-specific adoptive immunotherapy, remain unknown. In present study, we detected the dynamic expression of eight major checkpoint molecules (CTLA-4, PD-1, PD-L1, TIM- 3, CEACAM-1, LAG-3, TIGIT and BTLA) on CIK cells from NSCLC patients. The majority of these molecules, except BTLA, were sharply elevated during the early stage of CIK cell culture. Thereafter, PD-1 and TIGIT expressions decreased gradually towards the initial level (day 0). Moreover, CTLA-4 faded away during the later stage of CIK culture. LAG-3 expression decreased but was still significantly higher than the initial level. Of note, PD-L1 remained stably upregulated during CIK culture compared with PD-1, indicating that PD-L1 might act as an inhibitory molecule on CIK cells instead of PD-1. Furthermore, TIM-3 and CEACAM1 were strongly expressed simultaneously during long-term CIK culture and showed a significant and mutually positive correlation. BTLA displayed a distinct pattern, and its expression gradually decreased throughout the CIK culture. These observations suggested that CIK cells might be partly exhausted before clinical transfusion, characterized by the high expression of PD-L1, LAG-3, TIM- 3, and CEACAM-1 and the low expression of TIGIT, BTLA, PD-1, and CTLA-4 compared with initial culture. Our results imply that implementing combined treatment on CIK cells before transfusion via antibodies targeting PD-L1, LAG-3, TIM-3, and CEACAM-1 might improve the efficiency of CIK therapy for NSCLC patients.
Insights
Immune checkpoint molecules on cytokine-induced killer (CIK) cells in NSCLC patients were analyzed. Targeting PD-L1, LAG-3, TIM-3, and CEACAM-1 on CIK cells may enhance cancer immunotherapy effectiveness.
Area of Science:
- Immunology
- Cancer Biology
- Cell Therapy
Background:
- Immune checkpoints regulate T-cell function and are implicated in cancer immune evasion.
- Cytokine-induced killer (CIK) cells are a promising non-specific adoptive immunotherapy for cancer.
- The expression profile of immune checkpoint molecules on CIK cells remains largely unexplored.
Purpose of the Study:
- To investigate the dynamic expression of eight major immune checkpoint molecules on CIK cells from non-small cell lung cancer (NSCLC) patients during in vitro culture.
- To identify potential inhibitory molecules on CIK cells that could be targeted to improve immunotherapy efficacy.
Main Methods:
- Flow cytometry was used to detect the expression of CTLA-4, PD-1, PD-L1, TIM-3, CEACAM-1, LAG-3, TIGIT, and BTLA on CIK cells.
- CIK cells were cultured over time, and checkpoint molecule expression was analyzed at different time points.
- Correlative analysis was performed to assess the relationship between the expression of different checkpoint molecules.
Main Results:
- Most checkpoint molecules (except BTLA) were upregulated early in CIK culture, with PD-1 and TIGIT decreasing over time.
- PD-L1 remained stably upregulated, suggesting a potential inhibitory role on CIK cells.
- TIM-3 and CEACAM-1 showed strong, correlated expression during long-term culture, while BTLA expression decreased.
- CIK cells exhibited signs of exhaustion before transfusion, characterized by high PD-L1, LAG-3, TIM-3, CEACAM-1 and low TIGIT, BTLA, PD-1, CTLA-4.
Conclusions:
- CIK cells display dynamic immune checkpoint expression profiles during culture, potentially leading to exhaustion.
- Targeting PD-L1, LAG-3, TIM-3, and CEACAM-1 on CIK cells prior to transfusion may represent a viable strategy to enhance CIK-based immunotherapy for NSCLC.
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