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( A ) Scheme of the study flow and time points of blood withdrawal during the course of major surgery. ( B ) mRNA expression of IFNG (left), GZMB (middle) and PRF1 (right) in CTL (+CD3/CD28 dynabeads, stimulated for 18 h) relative to endogenous controls, n = 9/10/10 individual patients. ( C ) Flow cytometric analysis (FCA) of intracellular IFN-γ staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( D ) Quantification of IFN-γ secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h), n = 9 individual patients. ( E ) Representative impedance plot (left) and quantification of relative CTL cytotoxicity (right) using ECIS, n = 7 individual patients. ( F ) FCA of <t>CD38</t> staining and quantification in CTL, minimum = 28.6/32.9, 25%-percentile = 46.73/55.65, median = 59.65/67.05, 75%-percentile = 69.10/75.75, maximum = 75.10/80.60 (T1/T2), n = 10 individual patients. ( G ) Quantification of GZMB (left) and PRF1 (right) secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h) as measured by ELISA, n = 11/12/6 (GZMB) and 13/13/6 (PRF1) individual patients. ( H ) FCA of intracellular GZMB staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( I ) FCA of intracellular PRF1 staining and quantification in CTL, n = 9 individual patients. ( J ) FCA of CD107a staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 6 individual patients. If not stated otherwise, data are represented as mean ± SEM. P values as indicated, paired t -test or Wilcoxon matched-pairs signed-rank test, as appropriate. .
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( A ) Scheme of the study flow and time points of blood withdrawal during the course of major surgery. ( B ) mRNA expression of IFNG (left), GZMB (middle) and PRF1 (right) in CTL (+CD3/CD28 dynabeads, stimulated for 18 h) relative to endogenous controls, n = 9/10/10 individual patients. ( C ) Flow cytometric analysis (FCA) of intracellular IFN-γ staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( D ) Quantification of IFN-γ secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h), n = 9 individual patients. ( E ) Representative impedance plot (left) and quantification of relative CTL cytotoxicity (right) using ECIS, n = 7 individual patients. ( F ) FCA of <t>CD38</t> staining and quantification in CTL, minimum = 28.6/32.9, 25%-percentile = 46.73/55.65, median = 59.65/67.05, 75%-percentile = 69.10/75.75, maximum = 75.10/80.60 (T1/T2), n = 10 individual patients. ( G ) Quantification of GZMB (left) and PRF1 (right) secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h) as measured by ELISA, n = 11/12/6 (GZMB) and 13/13/6 (PRF1) individual patients. ( H ) FCA of intracellular GZMB staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( I ) FCA of intracellular PRF1 staining and quantification in CTL, n = 9 individual patients. ( J ) FCA of CD107a staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 6 individual patients. If not stated otherwise, data are represented as mean ± SEM. P values as indicated, paired t -test or Wilcoxon matched-pairs signed-rank test, as appropriate. .
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( A ) Scheme of the study flow and time points of blood withdrawal during the course of major surgery. ( B ) mRNA expression of IFNG (left), GZMB (middle) and PRF1 (right) in CTL (+CD3/CD28 dynabeads, stimulated for 18 h) relative to endogenous controls, n = 9/10/10 individual patients. ( C ) Flow cytometric analysis (FCA) of intracellular IFN-γ staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( D ) Quantification of IFN-γ secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h), n = 9 individual patients. ( E ) Representative impedance plot (left) and quantification of relative CTL cytotoxicity (right) using ECIS, n = 7 individual patients. ( F ) FCA of <t>CD38</t> staining and quantification in CTL, minimum = 28.6/32.9, 25%-percentile = 46.73/55.65, median = 59.65/67.05, 75%-percentile = 69.10/75.75, maximum = 75.10/80.60 (T1/T2), n = 10 individual patients. ( G ) Quantification of GZMB (left) and PRF1 (right) secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h) as measured by ELISA, n = 11/12/6 (GZMB) and 13/13/6 (PRF1) individual patients. ( H ) FCA of intracellular GZMB staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( I ) FCA of intracellular PRF1 staining and quantification in CTL, n = 9 individual patients. ( J ) FCA of CD107a staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 6 individual patients. If not stated otherwise, data are represented as mean ± SEM. P values as indicated, paired t -test or Wilcoxon matched-pairs signed-rank test, as appropriate. .
Anti Cd38 Pe, supplied by Miltenyi Biotec, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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( A ) Scheme of the study flow and time points of blood withdrawal during the course of major surgery. ( B ) mRNA expression of IFNG (left), GZMB (middle) and PRF1 (right) in CTL (+CD3/CD28 dynabeads, stimulated for 18 h) relative to endogenous controls, n = 9/10/10 individual patients. ( C ) Flow cytometric analysis (FCA) of intracellular IFN-γ staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( D ) Quantification of IFN-γ secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h), n = 9 individual patients. ( E ) Representative impedance plot (left) and quantification of relative CTL cytotoxicity (right) using ECIS, n = 7 individual patients. ( F ) FCA of CD38 staining and quantification in CTL, minimum = 28.6/32.9, 25%-percentile = 46.73/55.65, median = 59.65/67.05, 75%-percentile = 69.10/75.75, maximum = 75.10/80.60 (T1/T2), n = 10 individual patients. ( G ) Quantification of GZMB (left) and PRF1 (right) secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h) as measured by ELISA, n = 11/12/6 (GZMB) and 13/13/6 (PRF1) individual patients. ( H ) FCA of intracellular GZMB staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( I ) FCA of intracellular PRF1 staining and quantification in CTL, n = 9 individual patients. ( J ) FCA of CD107a staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 6 individual patients. If not stated otherwise, data are represented as mean ± SEM. P values as indicated, paired t -test or Wilcoxon matched-pairs signed-rank test, as appropriate. .

Journal: EMBO Molecular Medicine

Article Title: Mitochondrial damage drives T-cell immunometabolic paralysis after major surgery

doi: 10.1038/s44321-025-00324-1

Figure Lengend Snippet: ( A ) Scheme of the study flow and time points of blood withdrawal during the course of major surgery. ( B ) mRNA expression of IFNG (left), GZMB (middle) and PRF1 (right) in CTL (+CD3/CD28 dynabeads, stimulated for 18 h) relative to endogenous controls, n = 9/10/10 individual patients. ( C ) Flow cytometric analysis (FCA) of intracellular IFN-γ staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( D ) Quantification of IFN-γ secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h), n = 9 individual patients. ( E ) Representative impedance plot (left) and quantification of relative CTL cytotoxicity (right) using ECIS, n = 7 individual patients. ( F ) FCA of CD38 staining and quantification in CTL, minimum = 28.6/32.9, 25%-percentile = 46.73/55.65, median = 59.65/67.05, 75%-percentile = 69.10/75.75, maximum = 75.10/80.60 (T1/T2), n = 10 individual patients. ( G ) Quantification of GZMB (left) and PRF1 (right) secretion by CTL (+CD3/CD28 dynabeads, stimulated for 18 h) as measured by ELISA, n = 11/12/6 (GZMB) and 13/13/6 (PRF1) individual patients. ( H ) FCA of intracellular GZMB staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 9 individual patients. ( I ) FCA of intracellular PRF1 staining and quantification in CTL, n = 9 individual patients. ( J ) FCA of CD107a staining and quantification in CTL (+CD3/CD28 dynabeads, stimulated for 18 h; re-stimulation with PMA/Ionomycin for 4 h), n = 6 individual patients. If not stated otherwise, data are represented as mean ± SEM. P values as indicated, paired t -test or Wilcoxon matched-pairs signed-rank test, as appropriate. .

Article Snippet: CD38 APC , Miltenyi , 130-113-991.

Techniques: Expressing, Staining, Enzyme-linked Immunosorbent Assay