Subset-specific Metabolic Adaptation and Functional Remodeling of Gamma Delta T (γδ T) Cells in Critically Ill ICU Patients: A Single-center, Prospective, Observational Cohort Study.
试验速览
- 阶段
- 不适用
- 状态
- 招募中
- 发起方
- 入组人数
- 105
- 试验地点
- 1
- 主要终点
- Change in the Proportion of Cytotoxic γδT Cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) Among Total γδT Cells
研究概览
简要总结
This prospective observational cohort study investigates the subset-specific metabolic adaptation and functional remodeling of cytotoxic γδT cells in critically ill patients with and without sepsis. Emerging evidence indicates that γδT cells, as a bridge between innate and adaptive immunity, play a critical role in early anti-infection defense during sepsis. However, the functional status and underlying regulatory mechanisms of cytotoxic γδT cells in septic patients remain incompletely understood. Our preliminary single-cell transcriptomic analysis revealed that cytotoxic γδT cells from septic patients exhibit significant alterations in cytotoxicity-associated molecules (GZMB, PRF1, GNLY) and mitochondrial oxidative phosphorylation (OXPHOS) pathway genes, particularly COX6C, which correlates with cytotoxic effector molecule expression. This study aims to systematically characterize the proportion, cytotoxicity, and mitochondrial metabolic function of circulating cytotoxic γδT cells across three cohorts: healthy controls, critically ill non-septic patients, and critically ill septic patients. By integrating flow cytometry, mitochondrial function assays, and functional validation experiments, we seek to elucidate the role of COX6C-mediated mitochondrial metabolic abnormalities in cytotoxic γδT cell dysfunction, providing theoretical basis for understanding immune dysregulation in sepsis and identifying novel therapeutic targets.
研究设计
- 研究类型
- Observational
- 观察模型
- Cohort
- 时间视角
- Prospective
入排标准
- 年龄范围
- 18 Years 至 80 Years（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 是
入选标准
- •Healthy Control Group (NHC):
- •Age ≥ 18 years.
- •No acute or chronic major diseases.
- •Provide written informed consent.
- •Non-septic Critical Illness Group (CI-NS):
- •Age ≥ 18 years.
- •Admitted to the ICU and meeting the definition of critical illness.
- •Excluded from sepsis according to the Sepsis-3 criteria (infection + ΔSOFA ≥ 2 points).
- •Written informed consent provided by the participant or legally authorized representative.
- •Septic Critical Illness Group (CI-Sep):
- •Age ≥ 18 years.
- •Admitted to the ICU and meeting the Sepsis-3 criteria (infection + ΔSOFA ≥ 2 points).
- •Written informed consent provided by the participant or legally authorized representative.
排除标准
- •Age < 18 years.
- •Known immunodeficiency, HIV infection, active hematologic malignancy, or history of hematopoietic stem cell or solid organ transplantation within the past 3 months.
- •Receipt of T-cell-targeted immunosuppressive therapy (e.g., antithymocyte globulin, calcineurin inhibitors, mycophenolate mofetil, methotrexate, or high-dose corticosteroids >1 mg/kg/day prednisone equivalent) before ICU admission or within 24 hours after ICU admission.
- •Use of immune checkpoint inhibitors (e.g., anti-PD-1/PD-L1/CTLA-4 antibodies) within the past 6 weeks.
- •Expected ICU stay < 24 hours or imminent risk of death (moribund state).
- •Pregnancy or breastfeeding.
- •Active major bleeding.
- •Inability to obtain informed consent.
研究组 & 干预措施
Healthy Controls (NHCs)
Healthy volunteers without acute or chronic major diseases, aged ≥18 years, who have provided written informed consent. Participants undergo a single blood draw and rectal swab collection.
Critically Ill Non-Septic Patients (CI-NS)
Patients admitted to the ICU meeting the criteria for critical illness but without sepsis, as determined by the Sepsis-3 criteria (infection with ΔSOFA ≥ 2 points excluded). Participants are aged ≥18 years, with informed consent obtained from the patient or legally authorized representative. Blood and rectal swab samples are collected at three time points: within 24 hours of ICU admission (D0), Day 2 (D2), and Day 7 (D7). Clinical follow-up extends to 90 days.
Critically Ill Septic Patients (CI-Sep)
Patients admitted to the ICU meeting the Sepsis-3 criteria (infection with ΔSOFA ≥ 2 points). Participants are aged ≥18 years, with informed consent obtained from the patient or legally authorized representative. Blood and rectal swab samples are collected at three time points: within 24 hours of ICU admission (D0), Day 2 (D2), and Day 7 (D7). Clinical follow-up extends to 90 days.
结局指标
主要结局
Change in the Proportion of Cytotoxic γδT Cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) Among Total γδT Cells
时间窗: Day 2 post-ICU admission
Flow cytometric quantification of the frequency of Cytotoxic γδT cells, defined as TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺ cells, as a percentage of total γδT cells (TCRγδ⁺) in peripheral blood. This subset represents the cytotoxic effector population critical for early anti-infection defense.
次要结局
- Change in COX6C Expression in Cytotoxic γδT Cells(Day 2 post-ICU admission)
- Change in Cytotoxic Molecule Expression in Cytotoxic γδT Cells(Day 2 post-ICU admission)
- Change in Mitochondrial Mass in Cytotoxic γδT Cell(Day 2 post-ICU admission)
- Change in Mitochondrial Membrane Potential (TMRE) in Cytotoxic γδT Cells(Day 2 post-ICU admission)
- Change in Mitochondrial Membrane Potential (JC-1) in Cytotoxic γδT Cells(Day 2 post-ICU admission)
- Change in Oxygen Consumption Rate (OCR) in Cytotoxic γδT Cells(Day 2 post-ICU admission)
- Change in Extracellular Acidification Rate (ECAR) in Cytotoxic γδT Cells(Day 2 post-ICU admission)
- Change in Glycolytic Capacity of Cytotoxic γδT Cells at Serial Time Points(Day 0, Day 2, and Day 7 post-ICU admission)
- Change in Mitochondrial ROS Levels in Cytotoxic γδT Cells at Serial Time Points(Day 0, Day 2, and Day 7 post-ICU admission)
- Change in Cytotoxic γδT Cell Migratory Function(Day 2 post-ICU admission)
- Change in Total γδT Cell Proportion at Serial Time Points(Day 0, Day 2, and Day 7 post-ICU admission)
- Change in Cytotoxic γδT Cell Proportion at Serial Time Points(Day 0, Day 2, and Day 7 post-ICU admission)
- Change in COX6C Expression in Cytotoxic γδT Cells at Serial Time Points(Day 0, Day 2, and Day 7 post-ICU admission)
- Correlation Between Cytotoxic γδT Cell Glycolytic Capacity and APACHE II Score(Day 2 post-ICU admission)
- Correlation Between Cytotoxic γδT Cell Glycolytic Capacity and SOFA Score(Day 2 post-ICU admission)
- Correlation Between Cytotoxic γδT Cell Metabolic Parameters and vasopressor dose(Day 2 post-ICU admission)
- Correlation Between Cytotoxic γδT Cell Metabolic Parameters and plasma lactate levels(Day 2 post-ICU admission)
- Association Between Cytotoxic γδT Cell Glycolytic Capacity and 28-day all-cause mortality(Up to 90 days post-discharge)
- Association Between Cytotoxic γδT Cell Glycolytic Capacity and 90-day all-cause mortality(Up to 90 days post-discharge)
- Association Between Cytotoxic γδT Cell Glycolytic Capacity and in-hospital mortality(Up to 90 days post-discharge)
- Association Between Cytotoxic γδT Cell Glycolytic Capacity and ICU mortality(Up to 90 days post-discharge)
- Association Between Cytotoxic γδT Cell Glycolytic Capacity and hospital length of stay(Up to 90 days post-discharge)
研究者
Jiancheng Zhang
Principal Investigator
Union Hospital, Tongji Medical College, Huazhong University of Science and Technology