Metabolic and Functional Study of γδ T Cells in Critically Ill Patients
2026年9月16日 更新者：Jiancheng Zhang、Union Hospital, Tongji Medical College, Huazhong University of Science and Technology
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.
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.
調査の概要
研究の種類
観察的
入学 (推定)
105
連絡先と場所
このセクションには、調査を実施する担当者の連絡先の詳細と、この調査が実施されている場所に関する情報が記載されています。
研究場所
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Outside of the US
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Wuhan、Outside of the US、中国、430022
- 募集
- Department of Critical Care Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology
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コンタクト：
- Jiancheng Zhang, Dr.
- 電話番号：13554105815
- メール：zhjcheng1@126.com
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参加基準
研究者は、適格基準と呼ばれる特定の説明に適合する人を探します。これらの基準のいくつかの例は、人の一般的な健康状態または以前の治療です。
適格基準
就学可能な年齢
- 大人
- 高齢者
健康ボランティアの受け入れ
はい
サンプリング方法
非確率サンプル
調査対象母集団
The study population comprises three distinct groups: healthy volunteers, non-septic critically ill patients, and septic critically ill patients admitted to the intensive care unit.
Sepsis is defined according to the Sepsis-3 criteria (infection with an acute increase in SOFA score ≥2 points), while critical illness is based on standard ICU admission criteria.
All participants must be aged ≥18 years and provide written informed consent (from the participant or a legally authorized representative for those unable to consent).
Key exclusion criteria include known immunodeficiency, recent use of T-cell-targeted immunosuppressants or immune checkpoint inhibitors, expected ICU stay <24 hours, pregnancy, active major bleeding, or inability to obtain consent.
Consecutive enrollment will take place at the Department of Critical Care Medicine, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, with follow-up through 90 days after enrollment.
説明
Inclusion Criteria:
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.
Exclusion Criteria:
- 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.
研究計画
このセクションでは、研究がどのように設計され、研究が何を測定しているかなど、研究計画の詳細を提供します。
研究はどのように設計されていますか？
デザインの詳細
コホートと介入
グループ/コホート |
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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.
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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.
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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.
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この研究は何を測定していますか？
主要な結果の測定
結果測定 |
メジャーの説明 |
時間枠 |
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Change in the Proportion of Cytotoxic γδT Cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) Among Total γδT Cells
時間枠：Day 2 post-ICU admission
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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.
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Day 2 post-ICU admission
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二次結果の測定
結果測定 |
メジャーの説明 |
時間枠 |
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Change in COX6C Expression in Cytotoxic γδT Cells
時間枠：Day 2 post-ICU admission
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Measurement of COX6C (mitochondrial respiratory chain complex IV subunit) expression levels in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) at the mRNA level (by single-cell RNA sequencing) and protein level (by flow cytometry).
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Day 2 post-ICU admission
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Change in Cytotoxic Molecule Expression in Cytotoxic γδT Cells
時間枠：Day 2 post-ICU admission
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Flow cytometric quantification of the mean fluorescence intensity (MFI) of cytotoxic effector molecules including Granzyme B (GZMB), Perforin (PRF1), and Granulysin (GNLY) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺).
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Day 2 post-ICU admission
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Change in Mitochondrial Mass in Cytotoxic γδT Cell
時間枠：Day 2 post-ICU admission
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Flow cytometric quantification of mitochondrial mass using MitoTracker Green FM staining in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺).
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Day 2 post-ICU admission
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Change in Mitochondrial Membrane Potential (TMRE) in Cytotoxic γδT Cells
時間枠：Day 2 post-ICU admission
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Flow cytometric quantification of mitochondrial membrane potential using tetramethylrhodamine ethyl ester (TMRE) staining in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺).
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Day 2 post-ICU admission
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Change in Mitochondrial Membrane Potential (JC-1) in Cytotoxic γδT Cells
時間枠：Day 2 post-ICU admission
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Flow cytometric quantification of mitochondrial membrane potential using JC-1 dye (red/green fluorescence ratio) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺).
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Day 2 post-ICU admission
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Change in Oxygen Consumption Rate (OCR) in Cytotoxic γδT Cells
時間枠：Day 2 post-ICU admission
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Measurement of basal oxygen consumption rate reflecting oxidative phosphorylation (OXPHOS) capacity in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺).
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Day 2 post-ICU admission
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Change in Extracellular Acidification Rate (ECAR) in Cytotoxic γδT Cells
時間枠：Day 2 post-ICU admission
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Measurement of basal extracellular acidification rate reflecting glycolytic metabolic activity in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺).
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Day 2 post-ICU admission
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Change in Glycolytic Capacity of Cytotoxic γδT Cells at Serial Time Points
時間枠：Day 0, Day 2, and Day 7 post-ICU admission
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Longitudinal flow cytometric puromycin-incorporation assay (SCENITH method) to measure glycolytic capacity in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) at Day 0, Day 2, and Day 7 post-ICU admission, to track the temporal evolution of metabolic flexibility and stress-adaptive glycolytic reprogramming during the course of critical illness.
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Day 0, Day 2, and Day 7 post-ICU admission
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Change in Mitochondrial ROS Levels in Cytotoxic γδT Cells at Serial Time Points
時間枠：Day 0, Day 2, and Day 7 post-ICU admission
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Longitudinal flow cytometric quantification of mitochondrial reactive oxygen species (mitochondrial ROS) levels using MitoSOX Green staining in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) at Day 0, Day 2, and Day 7 post-ICU admission, reflecting oxidative stress dynamics.
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Day 0, Day 2, and Day 7 post-ICU admission
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Change in Cytotoxic γδT Cell Migratory Function
時間枠：Day 2 post-ICU admission
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Assessment of Cytotoxic γδT cell (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) chemotactic and migratory capacity using Transwell migration assay.
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Day 2 post-ICU admission
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Change in Total γδT Cell Proportion at Serial Time Points
時間枠：Day 0, Day 2, and Day 7 post-ICU admission
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Longitudinal flow cytometric quantification of the frequency of total γδT cells (TCRγδ⁺) as a percentage of total live CD3⁺ T cells in peripheral blood at Day 0, Day 2, and Day 7 post-ICU admission.
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Day 0, Day 2, and Day 7 post-ICU admission
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Change in Cytotoxic γδT Cell Proportion at Serial Time Points
時間枠：Day 0, Day 2, and Day 7 post-ICU admission
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Longitudinal flow cytometric quantification of the frequency of Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) as a percentage of total γδT cells at Day 0, Day 2, and Day 7 post-ICU admission, to track the temporal evolution of this cell subset.
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Day 0, Day 2, and Day 7 post-ICU admission
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Change in COX6C Expression in Cytotoxic γδT Cells at Serial Time Points
時間枠：Day 0, Day 2, and Day 7 post-ICU admission
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Longitudinal measurement of COX6C expression levels in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) at Day 0, Day 2, and Day 7 post-ICU admission, to assess the temporal dynamics of mitochondrial metabolic gene expression.
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Day 0, Day 2, and Day 7 post-ICU admission
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Correlation Between Cytotoxic γδT Cell Glycolytic Capacity and APACHE II Score
時間枠：Day 2 post-ICU admission
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Spearman rank correlation analysis between glycolytic capacity (measured by puromycin incorporation assay at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) with APACHE II (Acute Physiology and Chronic Health Evaluation II) score measured within the first 48 hours of ICU admission.
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Day 2 post-ICU admission
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Correlation Between Cytotoxic γδT Cell Glycolytic Capacity and SOFA Score
時間枠：Day 2 post-ICU admission
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Spearman rank correlation analysis between glycolytic capacity (measured by puromycin incorporation assay at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) with SOFA (Sequential Organ Failure Assessment) score measured within the first 48 hours of ICU admission.
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Day 2 post-ICU admission
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Correlation Between Cytotoxic γδT Cell Metabolic Parameters and vasopressor dose
時間枠：Day 2 post-ICU admission
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Correlation analysis between COX6C expression, glycolytic capacity, and mitochondrial ROS levels in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) with vasopressor dose (reported as norepinephrine equivalent dose, NEE).
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Day 2 post-ICU admission
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Correlation Between Cytotoxic γδT Cell Metabolic Parameters and plasma lactate levels
時間枠：Day 2 post-ICU admission
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Correlation analysis between COX6C expression, glycolytic capacity, and mitochondrial ROS levels in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) with plasma lactate levels.
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Day 2 post-ICU admission
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Association Between Cytotoxic γδT Cell Glycolytic Capacity and 28-day all-cause mortality
時間枠：Up to 90 days post-discharge
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Analysis of association between glycolytic capacity (measured at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) and 28-day all-cause mortality.
Patients will be stratified by vital status (survivors vs. non-survivors) and by hospital length of stay (above vs. below median) for comparative analysis.
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Up to 90 days post-discharge
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Association Between Cytotoxic γδT Cell Glycolytic Capacity and 90-day all-cause mortality
時間枠：Up to 90 days post-discharge
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Analysis of association between glycolytic capacity (measured at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) and 90-day all-cause mortality.
Patients will be stratified by vital status (survivors vs. non-survivors) and by hospital length of stay (above vs. below median) for comparative analysis.
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Up to 90 days post-discharge
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Association Between Cytotoxic γδT Cell Glycolytic Capacity and in-hospital mortality
時間枠：Up to 90 days post-discharge
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Analysis of association between glycolytic capacity (measured at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) and in-hospital mortality.
Patients will be stratified by vital status (survivors vs. non-survivors) and by hospital length of stay (above vs. below median) for comparative analysis.
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Up to 90 days post-discharge
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Association Between Cytotoxic γδT Cell Glycolytic Capacity and ICU mortality
時間枠：Up to 90 days post-discharge
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Analysis of association between glycolytic capacity (measured at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) and ICU mortality.
Patients will be stratified by vital status (survivors vs. non-survivors) and by hospital length of stay (above vs. below median) for comparative analysis.
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Up to 90 days post-discharge
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Association Between Cytotoxic γδT Cell Glycolytic Capacity and hospital length of stay
時間枠：Up to 90 days post-discharge
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Analysis of association between glycolytic capacity (measured at Day 2) in Cytotoxic γδT cells (TCRγδ⁺GZMB⁺PRF1⁺GNLY⁺) and hospital length of stay (days).
Patients will be stratified by vital status (survivors vs. non-survivors) and by hospital length of stay (above vs. below median) for comparative analysis.
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Up to 90 days post-discharge
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協力者と研究者
ここでは、この調査に関係する人々や組織を見つけることができます。
研究記録日
これらの日付は、ClinicalTrials.gov への研究記録と要約結果の提出の進捗状況を追跡します。研究記録と報告された結果は、国立医学図書館 (NLM) によって審査され、公開 Web サイトに掲載される前に、特定の品質管理基準を満たしていることが確認されます。
主要日程の研究
研究開始 (実際)
2026年6月21日
一次修了 (推定)
2027年4月15日
研究の完了 (推定)
2027年7月15日
試験登録日
最初に提出
2026年6月25日
QC基準を満たした最初の提出物
2026年7月1日
最初の投稿 (実際)
2026年7月2日
学習記録の更新
投稿された最後の更新 (実際)
2026年9月17日
QC基準を満たした最後の更新が送信されました
2026年9月16日
最終確認日
2026年9月1日
詳しくは
この情報は、Web サイト clinicaltrials.gov から変更なしで直接取得したものです。研究の詳細を変更、削除、または更新するリクエストがある場合は、register@clinicaltrials.gov。 までご連絡ください。 clinicaltrials.gov に変更が加えられるとすぐに、ウェブサイトでも自動的に更新されます。