Back to Search
Start Over
GFPuv-Expressing Recombinant Rickettsia typhi: a Useful Tool for the Study of Pathogenesis and CD8 + T Cell Immunology in R. typhi Infection
- Source :
- Infection and Immunity. 85
- Publication Year :
- 2017
- Publisher :
- American Society for Microbiology, 2017.
-
Abstract
- Rickettsia typhi is the causative agent of endemic typhus, a disease with increasing incidence worldwide that can be fatal. Because of its obligate intracellular life style, genetic manipulation of the pathogen is difficult. Nonetheless, in recent years, genetic manipulation tools have been successfully applied to rickettsiae. We describe here for the first time the transformation of R. typhi with the pRAM18dRGA plasmid that originally derives from Rickettsia amblyommatis and encodes the expression of GFPuv (green fluorescent protein with maximal fluorescence when excited by UV light). Transformed R. typhi ( R. typhi GFPuv ) bacteria are viable, replicate with kinetics similar to those of wild-type R. typhi in cell culture, and stably maintain the plasmid and GFPuv expression under antibiotic treatment in vitro and in vivo during infection of mice. CB17 SCID mice infected with R. typhi GFPuv succumb to the infection with kinetics similar to those for animals infected with wild-type R. typhi and develop comparable pathology and bacterial loads in the organs, demonstrating that the plasmid does not influence pathogenicity. In the spleen and liver of infected CB17 SCID mice, the bacteria are detectable by immunofluorescence microscopy in neutrophils and macrophages by histological staining. Finally, we show for the first time that transformed rickettsiae can be used for the detection of CD8 + T cell responses. GFP-specific restimulation of spleen cells from R. typhi GFPuv -infected BALB/c mice elicits gamma interferon (IFN-γ), tumor necrosis factor alpha (TNF-α), and interleukin 2 (IL-2) secretion by CD8 + T cells. Thus, R. typhi GFPuv bacteria are a novel, potent tool to study infection with the pathogen in vitro and in vivo and the immune response to these bacteria.
- Subjects :
- 0301 basic medicine
Interleukin 2
biology
T cell
030106 microbiology
Immunology
biology.organism_classification
Microbiology
Virology
03 medical and health sciences
030104 developmental biology
Infectious Diseases
Plasmid
medicine.anatomical_structure
Immune system
Rickettsia typhi
medicine
Cytotoxic T cell
Parasitology
Pathogen
CD8
medicine.drug
Subjects
Details
- ISSN :
- 10985522 and 00199567
- Volume :
- 85
- Database :
- OpenAIRE
- Journal :
- Infection and Immunity
- Accession number :
- edsair.doi...........9f08ad1a68d21dd0bbd170d951edca18