• Project number: F 2509
• Institution: Federal Institute for Occupational Safety and Health (BAuA)
• Status: Completed Project

Description:

Human breathing functions as the intake pathway for bioaerosol components into the respiratory tract. Although the respiratory tract possesses a large number of barriers that prevent the uptake of bioaerosol components, the lung can nevertheless serve as a portal of entry for them on account of its large surface area. When this occurs, contact with airborne biological agents may not only cause infections, but also have sensitising (allergenic) or toxic effects on individuals. 

Under the German Biological Agents Ordinance (Biostoffverordnung, BioStoffV), biological agents’ infectious potential is reflected in their classification into risk groups. The toxic effects of biological agents are considered independently of these classifications. However, there is a lack of reliable methods for their investigation, which should ideally be done without animal testing. Model studies of health-damaging effects on cultivated human lung cells conducted under laboratory conditions (in vitro studies) are a suitable means of identifying biological agents’ toxic characteristics.

Proteins perform extremely diverse functions within cells. Furthermore, if changes occur, for instance when they come into contact with biological agents, cells respond in ways that alter the functions, activity levels, or quantities of different proteins. Alterations of these kinds can be quantified by proteomic analyses, which examine the entirety of a cell’s proteins at a particular point in time. When a comparative proteomic analysis was carried out on lung cells, some of which had been in contact with biological toxins and some of which had not, this allowed specific markers to be identified that are indicative of toxic effects. Such markers can help researchers to reliably specify the toxic characteristics of biological toxins and so support the ongoing work being done by the Committee on Biological Agents (Ausschuss für Biologische Arbeitsstoffe, ABAS) as it considers how the toxicity of toxin-producing biological agents should be classified in the relevant Technical Rules for Biological Agents (Technische Regeln für Biologische Arbeitsstoffe, TRBAs) (TRBA 466, TRBA 460). 

The objective for this project was to establish the whole workflow for the proteomic analysis of bronchial epithelial lung cells. It involved the cultivation of human lung cells with or without contact with biological toxins, the extraction of the proteins from the cells, and the digestion of the proteins to peptides. The peptides were labelled with isobaric chemical tags, separated by means of liquid chromatography, and identified using mass spectrometry. Bioinformatic analyses were subsequently undertaken.