结核病患者中细胞因子诱导的记忆样自然杀伤细胞反应减弱。
Attenuated Cytokine-Induced Memory-Like Natural Killer Cell Responses to in Tuberculosis Patients.
作者信息
Liang Chen, Li Shanshan, Yuan Jinfeng, Song Yanhua, Ren Weicong, Wang Wei, Shang Yuanyuan, Tang Shenjie, Pang Yu
机构信息
Department of Bacteriology and Immunology, Beijing Key Laboratory on Drug-Resistant Tuberculosis Research, Beijing Chest Hospital, Capital Medical University /Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing, 101149, People's Republic of China.
Tuberculosis Clinical Medical Center, Beijing Chest Hospital, Capital Medical University /Beijing Tuberculosis and Thoracic Tumor Research Institute, Beijing, 101149, People's Republic of China.
出版信息
Infect Drug Resist. 2023 Apr 20;16:2349-2364. doi: 10.2147/IDR.S407742. eCollection 2023.
PURPOSE
This study aimed to investigate the phenotype, proliferation and functional alterations of cytokine-induced memory-like natural killer (CIML NK) cells from healthy subjects and TB patients, and assessed the efficacy of CIML NK cells in response to H37Rv-infected U937 cells in vitro.
METHODS
Fresh peripheral blood mononuclear cells (PBMCs) were isolated from healthy people and tuberculosis patients and activated for 16h using low-dose IL-15, or IL-12, IL-15, IL-18 combination or IL-12, IL-15, IL-18 and MTB H37Rv lysates, respectively, followed by low-dose IL-15 maintenance for another 7 days. Then, the PBMCs were co-cultured with K562 and H37Rv-infected U937, and the purified NK cells were co-cultured with H37Rv infected U937. The phenotype, proliferation and response function of CIML NK cells were assessed using flow cytometry. Finally, colony forming units were enumerated to confirm the survival of intracellular MTB.
RESULTS
CIML NK phenotypes from TB patients were similar to healthy controls. CIML NK cells undergo higher rates of proliferation after IL-12/15/18 pre-activation. Moreover, the poor expansion potential of CIML NK cells co-stimulated with MTB lysates. CIML NK cells from healthy individuals showed enhanced IFN-γ functional to H37Rv infected U937 cells, along with significantly enhanced killing of H37Rv. However, the CIML NK cells from TB patients show attenuated IFN-γ production and now enhanced the ability of killing intracellular MTB compared to those from healthy donors after co-cultured with H37Rv infected U937.
CONCLUSION
CIML NK cells from healthy individuals exist the increased ability of IFN-γ secretion and boosted anti-MTB activity in vitro, which from TB patients show impaired IFN-γ production and no enhanced anti-MTB activity compared to those from healthy donors. Additionally, we observe the poor expansion potential of CIML NK cells co-stimulated with antigens from MTB. These results open up new possibilities for NK cell-based anti-tuberculosis immunotherapeutic strategies.
目的
本研究旨在调查健康受试者和结核病患者中细胞因子诱导的记忆样自然杀伤（CIML NK）细胞的表型、增殖和功能改变，并评估CIML NK细胞在体外对感染H37Rv的U937细胞的反应效果。
方法
从健康人和结核病患者中分离新鲜外周血单个核细胞（PBMC），分别用低剂量IL-15、或IL-12、IL-15、IL-18组合或IL-12、IL-15、IL-18与结核分枝杆菌H37Rv裂解物激活16小时，随后用低剂量IL-15维持培养7天。然后，将PBMC与K562和感染H37Rv的U937共培养，将纯化的NK细胞与感染H37Rv的U937共培养。使用流式细胞术评估CIML NK细胞的表型、增殖和反应功能。最后，计数集落形成单位以确认细胞内结核分枝杆菌的存活情况。
结果
结核病患者的CIML NK表型与健康对照相似。IL-12/15/18预激活后，CIML NK细胞的增殖率更高。此外，与结核分枝杆菌裂解物共刺激的CIML NK细胞的扩增潜力较差。健康个体的CIML NK细胞对感染H37Rv的U937细胞表现出增强的IFN-γ功能，同时对H37Rv的杀伤显著增强。然而，与感染H37Rv的U937共培养后，结核病患者的CIML NK细胞显示IFN-γ产生减弱，且与健康供体来源的细胞相比，杀伤细胞内结核分枝杆菌的能力未增强。
结论
健康个体的CIML NK细胞在体外存在IFN-γ分泌能力增强和抗结核分枝杆菌活性增强的情况，而结核病患者的CIML NK细胞与健康供体来源的细胞相比，IFN-γ产生受损且抗结核分枝杆菌活性未增强。此外，我们观察到与结核分枝杆菌抗原共刺激的CIML NK细胞的扩增潜力较差。这些结果为基于NK细胞的抗结核免疫治疗策略开辟了新的可能性。
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