Engineering attenuated virulence of a Theileria annulata-infected macrophage.
Live attenuated vaccines are used to combat tropical theileriosis in North Africa, the Middle East, India, and China. The attenuation process is empirical and occurs only after many months, sometimes years, of in vitro culture of virulent clinical isolates. During this extensive culturing, attenuated lines lose their vaccine potential. To circumvent this we engineered the rapid ablation of the host cell transcription factor c-Jun, and within only 3 weeks the line engineered for loss of c-Jun activation displayed in vitro correlates of attenuation such as loss of adhesion, reduced MMP9 gelatinase activity, and diminished capacity to traverse Matrigel. Specific ablation of a single infected host cell virulence trait (c-Jun) induced a complete failure of Theileria annulata-transformed macrophages to disseminate, whereas virulent macrophages disseminated to the kidneys, spleen, and lungs of Rag2/γC mice. Thus, in this heterologous mouse model loss of c-Jun expression led to ablation of dissemination of T. annulata-infected and transformed macrophages. The generation of Theileria-infected macrophages genetically engineered for ablation of a specific host cell virulence trait now makes possible experimental vaccination of calves to address how loss of macrophage dissemination impacts the disease pathology of tropical theileriosis.
Laboratoire de Biologie Cellulaire Comparative des Apicomplexes, Faculté de Médicine, Université Paris Descartes - Sorbonne Paris Cité, Paris, France; Inserm U1016, Cnrs UMR8104, Cochin Institute, Paris, France.
Nadia
N
Echebli
Laboratoire de Parasitologie, Ecole Nationale de Médecine Vétérinaire, Sidi Thabet, Tunisia.
Moez
M
Mhadhbi
Laboratoire de Biologie Cellulaire Comparative des Apicomplexes, Faculté de Médicine, Université Paris Descartes - Sorbonne Paris Cité, Paris, France; Inserm U1016, Cnrs UMR8104, Cochin Institute, Paris, France.
Marie
M
Chaussepied
Laboratoire de Biologie Cellulaire Comparative des Apicomplexes, Faculté de Médicine, Université Paris Descartes - Sorbonne Paris Cité, Paris, France; Inserm U1016, Cnrs UMR8104, Cochin Institute, Paris, France.
Catherine
C
Vayssettes
Innate Immunity Unit, Department of Immunology, Pasteur Institute, Paris, France; Inserm U688, Pasteur Institute, Paris, France.
James P
JP
Di Santo
Laboratoire de Parasitologie, Ecole Nationale de Médecine Vétérinaire, Sidi Thabet, Tunisia; Institution de la Recherche et de l'Enseignement Supérieur Agricoles, Tunis, Tunisia.
Mohamed Aziz
MA
Darghouth
Laboratoire de Biologie Cellulaire Comparative des Apicomplexes, Faculté de Médicine, Université Paris Descartes - Sorbonne Paris Cité, Paris, France; Inserm U1016, Cnrs UMR8104, Cochin Institute, Paris, France.
Gordon
G
Langsley
D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D005260:Female; D005353:Fibronectins / Q000378:metabolism; D005818:Genetic Engineering; D007668:Kidney / Q000469:parasitology; D008168:Lung / Q000469:parasitology; D008264:Macrophages / Q000276:immunology* / Q000378:metabolism / Q000469:parasitology; D008297:Male; D051379:Mice; D009154:Mutation; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics*; D016052:Protozoan Vaccines / Q000276:immunology*; D013154:Spleen / Q000469:parasitology; D016796:Theileria annulata / Q000276:immunology / Q000472:pathogenicity*; D013801:Theileriasis / Q000517:prevention & control*; D014611:Vaccination; D014613:Vaccines, Attenuated / Q000276:immunology; D014774:Virulence
D005353:Fibronectins; D016755:Proto-Oncogene Proteins c-jun; D016052:Protozoan Vaccines; D014613:Vaccines, Attenuated