Research Article: The afc antifungal activity cluster, which is under tight regulatory control of ShvR, is essential for transition from intracellular persistence of Burkholderia cenocepacia to acute pro-inflammatory infection

Date Published: December 4, 2018

Publisher: Public Library of Science

Author(s): Margarida C. Gomes, Yara Tasrini, Sujatha Subramoni, Kirsty Agnoli, Joana R. Feliciano, Leo Eberl, Pamela Sokol, David O’Callaghan, Annette C. Vergunst, David Skurnik.

http://doi.org/10.1371/journal.ppat.1007473

Abstract

The opportunistic pathogen Burkholderia cenocepacia is particularly life-threatening for cystic fibrosis (CF) patients. Chronic lung infections with these bacteria can rapidly develop into fatal pulmonary necrosis and septicaemia. We have recently shown that macrophages are a critical site for replication of B. cenocepacia K56-2 and the induction of fatal pro-inflammatory responses using a zebrafish infection model. Here, we show that ShvR, a LysR-type transcriptional regulator that is important for biofilm formation, rough colony morphotype and inflammation in a rat lung infection model, is also required for the induction of fatal pro-inflammatory responses in zebrafish larvae. ShvR was not essential, however, for bacterial survival and replication in macrophages. Temporal, rhamnose-induced restoration of shvR expression in the shvR mutant during intramacrophage stages unequivocally demonstrated a key role for ShvR in transition from intracellular persistence to acute fatal pro-inflammatory disease. ShvR has been previously shown to tightly control the expression of the adjacent afc gene cluster, which specifies the synthesis of a lipopeptide with antifungal activity. Mutation of afcE, encoding an acyl-CoA dehydrogenase, has been shown to give similar phenotypes as the shvR mutant. We found that, like shvR, afcE is also critical for the switch from intracellular persistence to fatal infection in zebrafish. The closely related B. cenocepacia H111 has been shown to be less virulent than K56-2 in several infection models, including Galleria mellonella and rats. Interestingly, constitutive expression of shvR in H111 increased virulence in zebrafish larvae to almost K56-2 levels in a manner that absolutely required afc. These data confirm a critical role for afc in acute virulence caused by B. cenocepacia that depends on strain-specific regulatory control by ShvR. We propose that ShvR and AFC are important virulence factors of the more virulent Bcc species, either through pro-inflammatory effects of the lipopeptide AFC, or through AFC-dependent membrane properties.

Partial Text

B. cenocepacia belongs to the Burkholderia cepacia complex (Bcc), currently encompassing 21 officially named species [1,2]. These opportunistic bacteria are notorious pathogens of cystic fibrosis (CF) patients [3,4] and are emerging as the culprit of serious infections in non-CF conditions, both in and outside the hospital [5–8]. Infection of CF airways by bacteria belonging to the Bcc can be asymptomatic but often result in chronic infection with intermittent acute exacerbations resulting in progressive worsening of lung function [9,10]. B. cenocepacia is particularly associated with reduced survival and a high risk for development of unpredictable acute fatal necrotizing pneumonia and sepsis, termed “cepacia syndrome”. Bcc bacteria are well-known for their intrinsic resistance to stress conditions and antibiotics [11–13], complicating disease management and treatment strategies.

The LysR-type transcriptional regulator ShvR has been shown to tightly control the expression of an adjacent operon, called afc, involved in antifungal activity [25]. The zebrafish embryo model, in which persistent and acute infection caused by different Bcc strains can be studied in detail in the context of an innate immune response [29,30] was exploited to gain better insight into a role for ShvR and AfcE in virulence of B. cenocepacia K56-2. We found that while shvR and afcE are not required for bacterial persistence in the host, they are both essential for the induction of a robust fatal pro-inflammatory infection. Our study shows that ShvR and AfcE are key elements in the transition from an intracellular bacterial stage to disseminated pro-inflammatory infection in zebrafish larvae.

 

Source:

http://doi.org/10.1371/journal.ppat.1007473

 

0 0 vote
Article Rating
Subscribe
Notify of
guest
0 Comments
Inline Feedbacks
View all comments