Analysis of initial intracellular activation processes of immunocompetent cells after cardiopulmonary resuscitation (CPR)

Analysis of initial intracellular activation processes of immunocompetent cells after cardiopulmonary resuscitation (CPR) - REA

Status

Active, not recruiting

Phases

Unknown

Study type

Observational

Source

DRKS

Registry ID

DRKS00012940

Enrollment

Registered

2017-09-22

Start date

2003-02-24

Completion date

Unknown

Last updated

2025-04-07

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

Post-reuscitation syndrome I46.0

Interventions

Group 1: Observational study: OHCA survivors with sustained unconsciousness (GCS<8) after return of spontaneous circulation (ROSC) are included (for detailed inclusion criteria see study protocol). F

Eligibility

Sex/Gender

All

Age

18 Years to No maximum

Inclusion criteria

Inclusion criteria: Patients (=18 years) successfully resuscitated from OHCA with sustained unconsciousness (GCS<8) after ROSC are included.

Exclusion criteria

Exclusion criteria: Main exclusion criteria were acute infection, intracerebral hemorrhage, aortic dissection or pregnancy. Further, patients who die within the first 24h after CPR are excluded.

Design outcomes

Primary

Measure	Time frame
Primary outcome parameter: Analysis of initial intracellular activation processes of immunocompetent cells (monocytes/granulocytes) from whole blood samples (EDTA) and PAXgene whole blood samples after cardiopulmonary resuscitation (CPR). Methods: Blood sampling: To evaluate very early changes, samples were collected immediately after initiation of cardiopulmonary resuscitation by the attending emergency physician (central ambulance station/LMU Munich) and subsequently 6, 12, 24, 48 and 72h by one of the study`s investigators. Both EDTA whole blood samples and PAXgene whole blood tubes were drawn. Processing: EDTA blood samples: For isolation of cellular subsets magnetic microbeads (CD14/CD15) by Milteny Biotech (Bergisch Gladbach) were used. Afterwards samples were stored at -80°C. PAXgene tubes: Blood was collected in PAXgene tubes and incubated for 24h. Afterwards samples were stored at -80°C. RNA extraction For RNA extraction of monocytes and granulocytes RNA-Midi-Kit (Qiagen) were used. Total RNA of PAXgene was extracted using the PAXgene Blood RNA Kit (PreAnalytix, Hombrechtikon, Swiss) according to the manufacturer’s instructions. Further procedures of both sytems were equivalent: cDNA synthesis (Reverse Transkription, AMV, Firma Roche). Quantification of mRNA expression The quantitative analysis of target gene expression (TNFa, IL-1ß, IL-6, IL-8, IL-10, IL-1ra, PMN Elastase, CD 11a, Tyrosinkinase, HPRT) was performed on a LightCycler by real-time PCR using the Light Cycler Fast Start DNA Master Sybr Green I Kit according to the manufacture`s instructions (Roche, Mannheim, Germany). Standard and primer were designed by Search LC (Heidelberg, Germany; Light Cycler Primer Sets); transscription factors (NF-?B, AP-1) were quantified by „electric mobility shift assay“ (EMSA).	—

Measure

Time frame

Primary outcome parameter: Analysis of initial intracellular activation processes of immunocompetent cells (monocytes/granulocytes) from whole blood samples (EDTA) and PAXgene whole blood samples after cardiopulmonary resuscitation (CPR). Methods: Blood sampling: To evaluate very early changes, samples were collected immediately after initiation of cardiopulmonary resuscitation by the attending emergency physician (central ambulance station/LMU Munich) and subsequently 6, 12, 24, 48 and 72h by one of the study`s investigators. Both EDTA whole blood samples and PAXgene whole blood tubes were drawn. Processing: EDTA blood samples: For isolation of cellular subsets magnetic microbeads (CD14/CD15) by Milteny Biotech (Bergisch Gladbach) were used. Afterwards samples were stored at -80°C. PAXgene tubes: Blood was collected in PAXgene tubes and incubated for 24h. Afterwards samples were stored at -80°C. RNA extraction For RNA extraction of monocytes and granulocytes RNA-Midi-Kit (Qiagen) were used. Total RNA of PAXgene was extracted using the PAXgene Blood RNA Kit (PreAnalytix, Hombrechtikon, Swiss) according to the manufacturer’s instructions. Further procedures of both sytems were equivalent: cDNA synthesis (Reverse Transkription, AMV, Firma Roche). Quantification of mRNA expression The quantitative analysis of target gene expression (TNFa, IL-1ß, IL-6, IL-8, IL-10, IL-1ra, PMN Elastase, CD 11a, Tyrosinkinase, HPRT) was performed on a LightCycler by real-time PCR using the Light Cycler Fast Start DNA Master Sybr Green I Kit according to the manufacture`s instructions (Roche, Mannheim, Germany). Standard and primer were designed by Search LC (Heidelberg, Germany; Light Cycler Primer Sets); transscription factors (NF-?B, AP-1) were quantified by „electric mobility shift assay“ (EMSA).

—

Secondary

Measure	Time frame
Secondary outcome: Correlation of cytokine mRNA changed and distinct clinical parameters. Therefore, this study was further designed to 2) correlate cytokine levels with clinical outcome parameters and 3) evaluate the effect of TH on cytokine mRNA expression levels. Multiple organ failure (MOF) was detected using MOF score as described by Goris et al. and modified by Lefering and colleagues [1,2]. A MOF score = 4 was surmised as multiple organ failure. If clinical data sets were incomplete patients were excluded from this sub-analysis. Survival was defined as 30-days survival after cardiac arrest. Patients who did not survive the first 24h after resuscitation were excluded. Therapeutic hypothermia (TH) was not yet implemented in Resuscitation Guidelines at the time of sample collection and therefore not induced regularly. Our patients were treated in eight different hospitals and a standardized TH protocol was therefore unavailable. In order to reduce heterogeneity, patients were retrospectively divided into two sub-groups. Only patients treated according to the same TH protocol were included into the hypothermia group. Hypothermia was achieved by external cooling methods (blanket, ice packs) and associated with routine use of neuromuscular blocking agents. Targeted levels of hypothermia were 32-34°C and maintained for 24h. 1. Goris, R.J., Boekhorst, te, T.P., Nuytinck, J.K., Gimbrère, J.S.: Multiple-organ failure. Generalized autodestructive inflammation? Arch Surg. 120, 1109–1115 (1985). 2. Lefering, R., Goris, R.J.A., van Nieuwenhoven, E.J., Neugebauer, E.: Revision of the multiple organ failure score. Langenbecks Arch Surg. 387, 14–20 (2002).	—

Measure

Time frame

Secondary outcome: Correlation of cytokine mRNA changed and distinct clinical parameters. Therefore, this study was further designed to 2) correlate cytokine levels with clinical outcome parameters and 3) evaluate the effect of TH on cytokine mRNA expression levels. Multiple organ failure (MOF) was detected using MOF score as described by Goris et al. and modified by Lefering and colleagues [1,2]. A MOF score = 4 was surmised as multiple organ failure. If clinical data sets were incomplete patients were excluded from this sub-analysis. Survival was defined as 30-days survival after cardiac arrest. Patients who did not survive the first 24h after resuscitation were excluded. Therapeutic hypothermia (TH) was not yet implemented in Resuscitation Guidelines at the time of sample collection and therefore not induced regularly. Our patients were treated in eight different hospitals and a standardized TH protocol was therefore unavailable. In order to reduce heterogeneity, patients were retrospectively divided into two sub-groups. Only patients treated according to the same TH protocol were included into the hypothermia group. Hypothermia was achieved by external cooling methods (blanket, ice packs) and associated with routine use of neuromuscular blocking agents. Targeted levels of hypothermia were 32-34°C and maintained for 24h. 1. Goris, R.J., Boekhorst, te, T.P., Nuytinck, J.K., Gimbrère, J.S.: Multiple-organ failure. Generalized autodestructive inflammation? Arch Surg. 120, 1109–1115 (1985). 2. Lefering, R., Goris, R.J.A., van Nieuwenhoven, E.J., Neugebauer, E.: Revision of the multiple organ failure score. Langenbecks Arch Surg. 387, 14–20 (2002).

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Countries

Germany

Contacts

Public ContactMareen Braunstein

Klinik für Allgemeine, Unfall- und Wiederherstellungschirurgie der LMU München

Mareen.Braunstein@med.uni-muenchen.de089-4400-52511

Outcome results

None listed

Source: DRKS (via WHO ICTRP) · Data processed: Feb 27, 2026