Assessment of seven protocols to prepare Cutibacterium acnes bacterial lysates to measure its immunogenic potential and review of the literature. (June 2019)

• Record Type: Journal Article
• Authors: Dagnelie, Marie-Ange; Khammari, Amir; Dréno, Brigitte; Corvec, Stéphane
• Is Part Of: Anaerobe. Volume 57(2019)
• Page Start: 75
• Page End: 81
• Publication Date: 2019-06
• Subjects: Cutibacterium acnes -- Cell wall membrane -- Bacterial extract -- Innate immunity -- Skin explants -- Inflammation -- Biomarkers; Anaerobic infections -- Periodicals; Anaerobic bacteria -- Periodicals; Bacterial diseases -- Periodicals; Computer network resources; Anaerobic protozoa -- Periodicals; 579.3
• DOI: 10.1016/j.anaerobe.2019.03.019
• Languages: English
• ISSNs: 1075-9964

Abstract

Abstract: Seven protocols were tested to prepare cell wall extracts from live Cutibacterium acnes . Different parameters were modified: thawing/freezing and sonication/freezing cycles, to impact on mechanical degradation of the bacteria. Finally, the immunogenic potential of the extracts generated was evaluated by measuring IL-8 releases using an in vitro skin explants system. The aim of this article was to compare the existing protocols from the scientific literature, and also propose a standardized method developed in our facilities. Highlights: Developing methods to create membrane extracts is essential to enable antibiotics use in cell culture and improve results reproducibility. We propose a standardized method to create bacterial membrane extracts from a suspension of living C. acnes, using mainly mechanical disruptions. Three criteria were monitored to select one protocol: bacteria non-regeneration, lysates immunogenicity, and protein concentration. This article compares existing protocols and proposes a standardized procedure developed in our facilities.