GFPuv-expressing recombinant Rickettsia typhi: A useful tool for the study of pathogenesis and CD8+

Matthias Hauptmann, Nicole Burkhardt, Ulrike Munderloh, Svenja Kuehl, Ulricke Richardt, Susanne Krasemann, Kristin Hartmann, Till Krech, Bernhard Fleischer, Christian Keller, Anke Osterloh

Research output: Contribution to journalArticle

  • 1 Citations

Abstract

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. typhiGFPuv) 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. typhiGFPuv 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. typhiGFPuv-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. typhiGFPuv bacteria are a novel, potent tool to study infection with the pathogen in vitro and in vivo and the immune response to these bacteria.

Original languageEnglish (US)
Article numbere00156-17
JournalInfection and Immunity
Volume85
Issue number6
DOIs
StatePublished - Jun 1 2017

Fingerprint

Rickettsia typhi
Bacteria
Rickettsia
Plasmids
Infection
SCID Mice
Interferon-gamma
Spleen
Tumor Necrosis Factor-alpha
T-Lymphocytes
In Vitro Techniques
Endemic Flea-Borne Typhus
Bacterial Load
Ultraviolet Rays
Green Fluorescent Proteins
Fluorescence Microscopy
Interleukin-2
Virulence
Life Style
Neutrophils

Keywords

  • CD8 T cell response
  • GFPuv
  • Rickettsia typhi
  • Transformation

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Infectious Diseases

MeSH PubMed subject areas

  • Journal Article

Cite this

Hauptmann, M., Burkhardt, N., Munderloh, U., Kuehl, S., Richardt, U., Krasemann, S., ... Osterloh, A. (2017). GFPuv-expressing recombinant Rickettsia typhi: A useful tool for the study of pathogenesis and CD8+. Infection and Immunity, 85(6), [e00156-17]. DOI: 10.1128/IAI.00156-17

GFPuv-expressing recombinant Rickettsia typhi : A useful tool for the study of pathogenesis and CD8+. / Hauptmann, Matthias; Burkhardt, Nicole; Munderloh, Ulrike; Kuehl, Svenja; Richardt, Ulricke; Krasemann, Susanne; Hartmann, Kristin; Krech, Till; Fleischer, Bernhard; Keller, Christian; Osterloh, Anke.

In: Infection and Immunity, Vol. 85, No. 6, e00156-17, 01.06.2017.

Research output: Contribution to journalArticle

Hauptmann, M, Burkhardt, N, Munderloh, U, Kuehl, S, Richardt, U, Krasemann, S, Hartmann, K, Krech, T, Fleischer, B, Keller, C & Osterloh, A 2017, 'GFPuv-expressing recombinant Rickettsia typhi: A useful tool for the study of pathogenesis and CD8+' Infection and Immunity, vol 85, no. 6, e00156-17. DOI: 10.1128/IAI.00156-17
Hauptmann M, Burkhardt N, Munderloh U, Kuehl S, Richardt U, Krasemann S et al. GFPuv-expressing recombinant Rickettsia typhi: A useful tool for the study of pathogenesis and CD8+. Infection and Immunity. 2017 Jun 1;85(6). e00156-17. Available from, DOI: 10.1128/IAI.00156-17

Hauptmann, Matthias; Burkhardt, Nicole; Munderloh, Ulrike; Kuehl, Svenja; Richardt, Ulricke; Krasemann, Susanne; Hartmann, Kristin; Krech, Till; Fleischer, Bernhard; Keller, Christian; Osterloh, Anke / GFPuv-expressing recombinant Rickettsia typhi : A useful tool for the study of pathogenesis and CD8+.

In: Infection and Immunity, Vol. 85, No. 6, e00156-17, 01.06.2017.

Research output: Contribution to journalArticle

@article{3589cfd86498477d9588c5c0d6f4db34,
title = "GFPuv-expressing recombinant Rickettsia typhi: A useful tool for the study of pathogenesis and CD8+",
abstract = "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. typhiGFPuv) 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. typhiGFPuv 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. typhiGFPuv-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. typhiGFPuv bacteria are a novel, potent tool to study infection with the pathogen in vitro and in vivo and the immune response to these bacteria.",
keywords = "CD8 T cell response, GFPuv, Rickettsia typhi, Transformation",
author = "Matthias Hauptmann and Nicole Burkhardt and Ulrike Munderloh and Svenja Kuehl and Ulricke Richardt and Susanne Krasemann and Kristin Hartmann and Till Krech and Bernhard Fleischer and Christian Keller and Anke Osterloh",
year = "2017",
month = "6",
doi = "10.1128/IAI.00156-17",
volume = "85",
journal = "Infection and Immunity",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "6",

}

TY - JOUR

T1 - GFPuv-expressing recombinant Rickettsia typhi

T2 - Infection and Immunity

AU - Hauptmann,Matthias

AU - Burkhardt,Nicole

AU - Munderloh,Ulrike

AU - Kuehl,Svenja

AU - Richardt,Ulricke

AU - Krasemann,Susanne

AU - Hartmann,Kristin

AU - Krech,Till

AU - Fleischer,Bernhard

AU - Keller,Christian

AU - Osterloh,Anke

PY - 2017/6/1

Y1 - 2017/6/1

N2 - 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. typhiGFPuv) 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. typhiGFPuv 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. typhiGFPuv-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. typhiGFPuv bacteria are a novel, potent tool to study infection with the pathogen in vitro and in vivo and the immune response to these bacteria.

AB - 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. typhiGFPuv) 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. typhiGFPuv 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. typhiGFPuv-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. typhiGFPuv bacteria are a novel, potent tool to study infection with the pathogen in vitro and in vivo and the immune response to these bacteria.

KW - CD8 T cell response

KW - GFPuv

KW - Rickettsia typhi

KW - Transformation

UR - http://www.scopus.com/inward/record.url?scp=85019886624&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85019886624&partnerID=8YFLogxK

U2 - 10.1128/IAI.00156-17

DO - 10.1128/IAI.00156-17

M3 - Article

VL - 85

JO - Infection and Immunity

JF - Infection and Immunity

SN - 0019-9567

IS - 6

M1 - e00156-17

ER -