Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.

Pathogenic Rickettsia species target vascular endothelial cells and cause systemic vasculitis. As obligate intracellular bacterial pathogens, Rickettsia must secure nutritional resources within the cytoplasm of endothelial cells while simultaneously subverting the innate immune defense system. With...

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Main Authors: Smruti Mishra, Luke Helminiak, Hwan Keun Kim
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2025-06-01
Series:PLoS Pathogens
Online Access:https://doi.org/10.1371/journal.ppat.1013277
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author Smruti Mishra
Luke Helminiak
Hwan Keun Kim
author_facet Smruti Mishra
Luke Helminiak
Hwan Keun Kim
author_sort Smruti Mishra
collection DOAJ
description Pathogenic Rickettsia species target vascular endothelial cells and cause systemic vasculitis. As obligate intracellular bacterial pathogens, Rickettsia must secure nutritional resources within the cytoplasm of endothelial cells while simultaneously subverting the innate immune defense system. With advances in rickettsial and host genetics, recent studies have identified novel molecular mechanisms involved in the complex interactions between Rickettsia and endothelial cells. However, it remains unclear how Rickettsia shields pathogen-derived immune stimulants, such as lipopolysaccharides (LPS) and peptidoglycan fragments, from immune recognition during intracellular replication. Prior work described two Rickettsia conorii variants with kkaebi transposon insertions in the polysaccharide synthesis operon (pso). Biochemical and immunological analyses revealed that pso is responsible for the biosynthesis of O-antigen (O-Ag) and the proper assembly of surface proteins. In the present work, we document that pso variant HK2 exhibits reduced capacities to adhere to and invade microvascular endothelial cells. Despite the low intracellular abundance, HK2 induced significantly higher levels of proinflammatory cytokines and chemokines, leading to premature cell death. Notably, HK2 exhibited defective intracellular survival in bone marrow-derived macrophages. This inability to dampen endothelial cell-mediated immune stimulation and resist macrophage-induced bactericidal activities resulted in the rapid elimination of viable Rickettsia in the mouse model of spotted fever. Further, when tested as a live-attenuated vaccine, HK2 elicited robust protective immunity against lethal spotted fever pathogenesis. Our work highlights the crucial role of pso in enabling Rickettsia to evade immune surveillance during intracellular replication within endothelial cells, ultimately delaying pathogen-induced programmed cell death and escaping immune defense mechanisms.
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spelling doaj-art-ad77c3f80c1246dc9fa1c296b4afe33f2025-08-20T02:46:44ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742025-06-01216e101327710.1371/journal.ppat.1013277Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.Smruti MishraLuke HelminiakHwan Keun KimPathogenic Rickettsia species target vascular endothelial cells and cause systemic vasculitis. As obligate intracellular bacterial pathogens, Rickettsia must secure nutritional resources within the cytoplasm of endothelial cells while simultaneously subverting the innate immune defense system. With advances in rickettsial and host genetics, recent studies have identified novel molecular mechanisms involved in the complex interactions between Rickettsia and endothelial cells. However, it remains unclear how Rickettsia shields pathogen-derived immune stimulants, such as lipopolysaccharides (LPS) and peptidoglycan fragments, from immune recognition during intracellular replication. Prior work described two Rickettsia conorii variants with kkaebi transposon insertions in the polysaccharide synthesis operon (pso). Biochemical and immunological analyses revealed that pso is responsible for the biosynthesis of O-antigen (O-Ag) and the proper assembly of surface proteins. In the present work, we document that pso variant HK2 exhibits reduced capacities to adhere to and invade microvascular endothelial cells. Despite the low intracellular abundance, HK2 induced significantly higher levels of proinflammatory cytokines and chemokines, leading to premature cell death. Notably, HK2 exhibited defective intracellular survival in bone marrow-derived macrophages. This inability to dampen endothelial cell-mediated immune stimulation and resist macrophage-induced bactericidal activities resulted in the rapid elimination of viable Rickettsia in the mouse model of spotted fever. Further, when tested as a live-attenuated vaccine, HK2 elicited robust protective immunity against lethal spotted fever pathogenesis. Our work highlights the crucial role of pso in enabling Rickettsia to evade immune surveillance during intracellular replication within endothelial cells, ultimately delaying pathogen-induced programmed cell death and escaping immune defense mechanisms.https://doi.org/10.1371/journal.ppat.1013277
spellingShingle Smruti Mishra
Luke Helminiak
Hwan Keun Kim
Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.
PLoS Pathogens
title Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.
title_full Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.
title_fullStr Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.
title_full_unstemmed Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.
title_short Polysaccharide synthesis operon modulates Rickettsia-endothelial cell interactions.
title_sort polysaccharide synthesis operon modulates rickettsia endothelial cell interactions
url https://doi.org/10.1371/journal.ppat.1013277
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AT lukehelminiak polysaccharidesynthesisoperonmodulatesrickettsiaendothelialcellinteractions
AT hwankeunkim polysaccharidesynthesisoperonmodulatesrickettsiaendothelialcellinteractions