Opsonisation improves duel proteomic analysis of Burkholderia cenocepacia infections. -DIA infection +/- Opsonisation of T6SS and TecA

ABSTRACT: The opportunistic Cystic Fibrosis (CF) pathogen Burkholderia cenocepacia is associated with severe lung infections. Within the CF lung, macrophages are both a crucial reservoir and a key mediator of the intense inflammatory response characteristic of B. cenocepacia infections. Despite the importance of macrophages for B. cenocepacia pathogenesis, there is a paucity of insight into how B. cenocepacia internalisation and replication impacts both the host and B. cenocepacia proteomes. A key limitation for understanding the proteomic changes during intracellular replication of B. cenocepacia has been the low infectivity of this pathogen, resulting in the generation of mixed cell populations dominated by uninfected cells within in vitro models. Using antibody-mediated opsonization, we show that by improving the efficiency and uniformity of B. cenocepacia internalization this enhances dual proteomic analysis. Comparing opsonized and non-opsonized infections, we show that changes in both host and internalized B. cenocepacia can be assessed in a single experimental framework. At the proteome level, opsonization dramatically improves the detection of protein changes, enhancing the detection of pro-inflammatory signaling and macrophage activation markers. Utilizing this dual proteomic approach, we assess the impact of the B. cenocepacia type 6 secretion system (T6SS) and the T6SS effector TecA at 3 and 24 hours post infection demonstrating that the presence/activities of the T6SS or TecA do not greatly modulate the proinflammatory response of THP-1 cells. Thus, this work demonstrates a simple means for enhancing proteomic analysis of B. cenocepacia infections enabling dual proteomic studies.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Nichollas Scott  

LAB HEAD: Nichollas E. Scott

PROVIDER: PXD073205 | Pride | 2026-05-13

REPOSITORIES: pride