Extracellular proteasome in the alveolar space: characterisation and function under physiological and pathophysiological condition

ISRCTN	ISRCTN20709647
DOI	https://doi.org/10.1186/ISRCTN20709647
Protocol serial number	N/A
Sponsor	German Research Council (Deutsche Forschungsgemeinschaft [DFG]) (Germany)
Funder	German Research Council (Deutsche Forschungsgemeinschaft [DFG]) (Germany) (ref: PE 301/4-1) - submitted

Submission date: 15/09/2007
Registration date: 19/10/2007
Last edited: 18/09/2017

Recruitment status: No longer recruiting
Overall study status: Completed
Condition category: Respiratory

Prospectively registered

Protocol

Statistical analysis plan

Results

Individual participant data

Plain English summary of protocol

Not provided at time of registration

Contact information

Dr Stephan Urs Sixt
Scientific

Klinik fur Anasthesiologie und Intensivmedizin
Hufelandstrasse 55
Essen
45122
Germany

Study information

Primary study design	Observational
Study design	This study is an observational, single-centre study. The aim of this study is the description and characterisation of the extracellular alveolar proteasome in healthy and sick subjects.
Secondary study design	Case-control study
Scientific title	Extracellular proteasome in the alveolar space: characterisation and function under physiological and pathophysiological condition: an observational study
Study acronym	ALIHD
Study objectives	We hypothesised that 20S proteasome is present and functional in the extracellular alveolar space of healthy subjects and patients with Acute Respiratory Distress Syndrome (ARDS), sarcoidosis and in the alveolar space after lung transplantation in humans.
Ethics approval(s)	Ethics approval received from the local ethics committee (Vorsitzender der Ethikkommission, Institut für Pharmakologie, Universitätsklinikum Essen) on the 9th December 2002 (ref: 01-97-1697).
Health condition(s) or problem(s) studied	Extracellular proteasome in the alveolar space
Intervention	Healthy caucasians subjects and patients with ARDS, sarcoidosis and patients after lung transplantation were included after approval of the local ethics committee and informed written consent. Observational trial: Proteasomal activity will be measured in Broncho-Alveolar Lavage (BAL) supernatant from healthy subjects, patients with acute respiratory distress syndrome and lung transplantation using specific proteasomal fluorogenic substrates and I125 albumin, with and without specific proteasome inhibitors. After that the different enzyme activities of the study groups were compared with healthy subjects. Furthermore, gel filtration, western blot technique, Enzyme-Linked Immuno-Sorbent Assay (ELISA) and mass spectrometry, were applied for proteasome characterisation in the different study groups.
Intervention type	Other
Primary outcome measure(s)	1. Enzyme activity using specific proteasomal fluorogenic substrates and I125 albumin, with and without specific proteasome inhibitors 2. Detection of 20S proteasome with polyclonal antibody (western-blot) 3. Detection of immunoproteasome with LMP2 and LMP7 antibody (western-blot) 4. Gel filtration to evaluate the molecular weight of the extracellular proteasome in the bronchoalveolar space 5. Mass spectrometry of the gel filtration revealed hydrolysing activity and ELISA technique for the amount of extracellular, alveolar proteasome
Key secondary outcome measure(s)	No secondary outcome measures
Completion date	01/01/2008

Eligibility

Participant type(s)	Patient
Age group	Adult
Lower age limit	18 Years
Upper age limit	70 Years
Sex	All
Target sample size at registration	300
Key inclusion criteria	1. Acute respiratory distress syndrome: 1.1. Timing: acute onset 1.2. Oxygenation: Partial Pressure of Oxygen in Arterial Blood (PaO2)/Fraction of Inspired Oxygen (FiO2) ratio less than 200 mmHg (regardless of Positive End Expiratory Pressure [PEEP]) 1.3. Chest radiograph: bilateral infiltrates seen on frontal chest radiograph 1.4. Pulmonary Artery Wedge (PAW): less than 18 mmHg when measured or no clinical evidence of left atrial hypertension 2. Sarcoidosis 3. Lung transplantation: patient after lung transplantation were lavaged in a routine diagnostic, for the screening of organ rejection and infection 4. Aged 18 to 70 years, both genders
Key exclusion criteria	1. Acute respiratory distress syndrome: contraindication against bronchoscopy 2. Sarcoidosis: contraindication against bronchoscopy 3. Lung transplantation: contraindication against bronchoscopy 4. Healthy subjects: 4.1. Contraindication against bronchoscopy 4.2. Any lung diseases 4.3. Infection 4.4. Sepsis 4.5. Coronary heart disease
Date of first enrolment	01/01/2003
Date of final enrolment	01/01/2008

Locations

Countries of recruitment

Germany

Study participating centre

Klinik fur Anasthesiologie und Intensivmedizin

Essen
45122
Germany

Results and Publications

Individual participant data (IPD) Intention to share	No
IPD sharing plan summary	Not provided at time of registration
IPD sharing plan

Study outputs

Output type	Details	Date created	Date added	Peer reviewed?	Patient-facing?
Results article	additional results	01/10/2014		Yes	No
Interim results article	initial findings	01/05/2007		Yes	No

Editorial Notes

18/09/2017: Publication reference added.