Establishment of an ELISA for the Recognition of Procalcitonin Variants in Patients With Hyperprocalcitonemia.

Establishment of an Enzyme-linked Immunosorbent Assay for the Recognition of Procalcitonin Variants and Characterization of Procalcitonin Variants in Patients With Hyperprocalcitonemia.

Procalcitonin is a protein consisting of 116 amino-acids which can rapidly rise under inflammatory conditions and sepsis. More than 20 years ago it has been shown that dipeptidylpeptidase-4 (DPP-4) cleaves procalcitonin from the n-terminus, resulting in a truncated procalcitonin-variant which consists of 114 aminoacids. Within their workgroup the investigators found that the truncated procalcitonin-variant had deleterious effects on vascular integrity during sepsis in mice. However, it is unknown if this applies also in humans. By using an ELISA-assay the investigators want to examine the ratio between native and truncated human procalcitonin during diseases accompanied with hyperprocalcitoninemia and correlate the results with clinical data.

Study Overview

• Status: Completed
• Conditions: Sepsis; Pre-Eclampsia; Adiposity; Microscopic Polyangiitis; Granulomatosis With Polyangiitis; SIRS; Procalcitonin
• Intervention / Treatment: Diagnostic test: Procalcitonin-variants ELISA-Assay

Detailed Description

Procalcitonin is a protein consisting of 116 amino-acids which can rapidly rise under inflammatory conditions and sepsis. More than 20 years ago it has been shown that dipeptidylpeptidase-4 (DPP-4) cleaves procalcitonin from the n-terminus, resulting in a truncated procalcitonin-variant which consists of 114 aminoacids.

Within their workgroup the investigators found that the truncated procalcitonin-variant had deleterious effects on vascular integrity during sepsis in mice: They observed that binding of truncated procalcitonin to the CRLR/RAMP1-receptor on vascular endothelium lead to phosphorylation and destruction of VE-cadherin, an essential part of adherens junctions. Consequently, paracellular leakage of proteins and fluid from blood vessels developed.

It is unknown if these effects also apply to humans. By using an ELISA-assay the investigators want to examine the ratio between native and truncated human procalcitonin during diseases accompanied with hyperprocalcitoninemia and correlate the results with clinical data. Futhermore, they want to examine if the procalcitonin-variants have influence on cytokine levels and surface antigens on immune cells by performing multiplex immunoassays and FACS-analysis.

• Study Type: Observational
• Enrollment (Actual): 30

Contacts and Locations

Study Locations

• Germany
  • North Rhine-Westphalia
    • Münster, North Rhine-Westphalia, Germany, 48147
      • University Hospital Münster

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years and older (Adult, Older Adult)
• Accepts Healthy Volunteers: N/A

• Sampling Method: Non-Probability Sample

Study Population

Patients from the University Hospital Münster who suffer from different diseases which are accompanied by hyperprocalcitoninemia.

Description

Inclusion Criteria:

• Age >18
• Patients with diagnosis...
• Sepsis or,
• SIRS after cardiothoracic surgery or,
• adipositas or,
• granulomatosis with polyangiitis/microscopic polyangiitis or,
• pre-eclampsia
• healthy control subjects
• written informed consent

Exclusion Criteria:

• participation in an interventional study trial within the last 3 months
• relationship to study investigator

Study Plan

How is the study designed?

Design Details

• Observational Models: Case-Control
• Time Perspectives: Cross-Sectional

Number of groups / cohorts

6

Cohorts and Interventions

Group / Cohort	Intervention / Treatment
Sepsis; Withdrawal of 3 blood collection tubes for ELISA-measurements at a single time during hyperprocalcitoninemia.	Diagnostic test: Procalcitonin-variants ELISA-Assay; Observational study measuring procalcitonin-variants in different patient collectives by obtaining 3 blood collection tubes per patient.
SIRS; Withdrawal of 3 blood collection tubes for ELISA-measurements at a single time during hyperprocalcitoninemia.	Diagnostic test: Procalcitonin-variants ELISA-Assay; Observational study measuring procalcitonin-variants in different patient collectives by obtaining 3 blood collection tubes per patient.
Adiposity; Withdrawal of 3 blood collection tubes for ELISA-measurements at a single time during hyperprocalcitoninemia.	Diagnostic test: Procalcitonin-variants ELISA-Assay; Observational study measuring procalcitonin-variants in different patient collectives by obtaining 3 blood collection tubes per patient.
Granulomatosis with polyangiitis / microscopic polyangiitis; Withdrawal of 3 blood collection tubes for ELISA-measurements at a single time during hyperprocalcitoninemia.	Diagnostic test: Procalcitonin-variants ELISA-Assay; Observational study measuring procalcitonin-variants in different patient collectives by obtaining 3 blood collection tubes per patient.
Pre-eclampsia; Withdrawal of 3 blood collection tubes for ELISA-measurements at a single time during hyperprocalcitoninemia.	Diagnostic test: Procalcitonin-variants ELISA-Assay; Observational study measuring procalcitonin-variants in different patient collectives by obtaining 3 blood collection tubes per patient.
Healthy controls; Withdrawal of 3 blood collection tubes for ELISA-measurements at a single time during hyperprocalcitoninemia.	Diagnostic test: Procalcitonin-variants ELISA-Assay; Observational study measuring procalcitonin-variants in different patient collectives by obtaining 3 blood collection tubes per patient.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Ratio between native and truncated procalcitonin during different conditions of hyperprocalcitoninemia	Measurement performed by using ELISA-assay.	Blood withdrawal takes approximately 5 minutes per patient

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
DPP-4-activity	Measurement performed by using a commercial DPP4-ELISA-kit.	Blood withdrawal takes approximately 5 minutes per patient
Proinflammatory cytokines	Measurement performed by using a commercial multiplex immunoassay kit.	Blood withdrawal takes approximately 5 minutes per patient
Immun cell surface-antigens	Measurement performed by using fluorescence-acivated cell sorting (FACS).	Blood withdrawal takes approximately 5 minutes per patient

Collaborators and Investigators

• Sponsor: University Hospital Muenster

Publications and helpful links

General Publications

• Wrenger S, Kahne T, Bohuon C, Weglohner W, Ansorge S, Reinhold D. Amino-terminal truncation of procalcitonin, a marker for systemic bacterial infections, by dipeptidyl peptidase IV (DP IV). FEBS Lett. 2000 Jan 21;466(1):155-9. doi: 10.1016/s0014-5793(99)01779-2.
• Brabenec L, Muller M, Hellenthal KEM, Karsten OS, Pryvalov H, Otto M, Holthenrich A, Matos ALL, Weiss R, Kintrup S, Hessler M, Dell'Aquila A, Thomas K, Nass J, Margraf A, Nottebaum AF, Rossaint J, Zarbock A, Vestweber D, Gerke V, Wagner NM. Targeting Procalcitonin Protects Vascular Barrier Integrity. Am J Respir Crit Care Med. 2022 Aug 15;206(4):488-500. doi: 10.1164/rccm.202201-0054OC.
• Weglohner W, Struck J, Fischer-Schulz C, Morgenthaler NG, Otto A, Bohuon C, Bergmann A. Isolation and characterization of serum procalcitonin from patients with sepsis. Peptides. 2001 Dec;22(12):2099-103. doi: 10.1016/s0196-9781(01)00541-1.

Study record dates

Study Major Dates

• Study Start (Actual): February 1, 2022
• Primary Completion (Actual): November 1, 2022
• Study Completion (Actual): November 1, 2022

Study Registration Dates

• First Submitted: January 19, 2023
• First Submitted That Met QC Criteria: January 19, 2023
• First Posted (Actual): January 30, 2023

Study Record Updates

• Last Update Posted (Estimated): June 4, 2024
• Last Update Submitted That Met QC Criteria: June 2, 2024
• Last Verified: June 1, 2024

More Information

Terms related to this study

• Additional Relevant MeSH Terms: Cardiovascular Diseases; Vascular Diseases; Cerebrovascular Disorders; Brain Diseases; Central Nervous System Diseases; Nervous System Diseases; Skin Diseases; Respiratory Tract Diseases; Immune System Diseases; Autoimmune Diseases; Lung Diseases; Vasculitis; Skin Diseases, Vascular; Pregnancy Complications; Lung Diseases, Interstitial; Hypertension, Pregnancy-Induced; Cerebral Small Vessel Diseases; Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis; Female Urogenital Diseases and Pregnancy Complications; Urogenital Diseases; Eclampsia; Pre-Eclampsia; Granulomatosis with Polyangiitis; Microscopic Polyangiitis; Systemic Vasculitis
• Other Study ID Numbers: 10-AnIt-19

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No