Orientia tsutsugamushi selectively stimulates the C-type lectin receptor Mincle and type 1-skewed proinflammatory immune responses.

James Fisher,Lynn Soong,Yuejin Liang,Holly Rosenzweig,Galen Card,Brandon Trent,J Stephen Dumler

doi:10.1371/journal.ppat.1009782

Abstract

Orientia tsutsugamushi is an obligately intracellular bacterium and the etiological agent of scrub typhus. The lung is a major target organ of infection, displaying type 1-skewed proinflammatory responses. Lung injury and acute respiratory distress syndrome are common complications of severe scrub typhus; yet, their underlying mechanisms remain unclear. In this study, we investigated whether the C-type lectin receptor (CLR) Mincle contributes to immune recognition and dysregulation. Following lethal infection in mice, we performed pulmonary differential expression analysis with NanoString. Of 671 genes examined, we found 312 significantly expressed genes at the terminal phase of disease. Mincle (Clec4e) was among the top 5 greatest up-regulated genes, accompanied with its signaling partners, type 1-skewing chemokines (Cxcr3, Ccr5, and their ligands), as well as Il27. To validate the role of Mincle in scrub typhus, we exposed murine bone marrow-derived macrophages (MΦ) to live or inactivated O. tsutsugamushi and analyzed a panel of CLRs and proinflammatory markers via qRT-PCR. We found that while heat-killed bacteria stimulated transitory Mincle expression, live bacteria generated a robust response in MΦ, which was validated by indirect immunofluorescence and western blot. Notably, infection had limited impact on other tested CLRs or TLRs. Sustained proinflammatory gene expression in MΦ (Cxcl9, Ccl2, Ccl5, Nos2, Il27) was induced by live, but not inactivated, bacteria; infected Mincle-/- MΦ significantly reduced proinflammatory responses compared with WT cells. Together, this study provides the first evidence for a selective expression of Mincle in sensing O. tsutsugamushi and suggests a potential role of Mincle- and IL-27-related pathways in host responses to severe infection. Additionally, it provides novel insight into innate immune recognition of this poorly studied bacterium.

Highlights

Scrub typhus is a vector-borne febrile illness caused by the obligately intracellular bacterium, Orientia tsutsugamushi
Scrub typhus is a life-threatening disease caused by the bacterium Orientia tsutsugamushi
It is thought that dysregulated immune responses contribute to the development of severe disease; it remains unknown as to how the immune system senses and responds to O. tsutsugamushi

Summary

Introduction

Scrub typhus is a vector-borne febrile illness caused by the obligately intracellular bacterium, Orientia tsutsugamushi. Recent reports have indicated the presence of scrub typhus in areas previously thought free of the disease, including South America and Africa [2,3]. The lung is a major target organ of infection, and mild interstitial pneumonia predominates in self-limiting or appropriately treated cases [1]. If left untreated, disease may progress to severe lung damage and acute respiratory distress syndrome in up to 25% of cases [1,4]. While the facets underpinning progression from mild to severe disease remain ill-defined, both bacterial and host factors are speculated to play major roles

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