title: In vivo near-infrared fluorescence imaging of Leishmania amazonensis expressing infrared fluorescence protein (iRFP) for real-time monitoring of cutaneous leishmaniasis in mice.
abstract: The use of Leishmania amazonensis-infected BALB/c mice is an important model for the study of experimental cutaneous leishmaniasis. Here we report the development of a non-invasive method to directly evaluate and measure parasite burden during the course of the infection, based on the near-infrared fluorescence detection of a recombinant L. amazonensis strain. So, we generated a L. amazonensis strain that stably expresses the near-infrared protein (iRFP) gene and compared the maintenance of its vitro and in vivo characteristics, such as fitness, pathogenicity and fluorescence emission. After that, we followed the disease development, as well as the parasite burden in BALB/c mice footpads infected with L. amazonensis-iRFP, by using an in vivo near-infrared fluorescence scanner. In vitro results showed a linear correlation between the fluorescence emission and the number of parasites. The in vivo study showed that the use of iRFP-transfected L. amazonensis enables the monitoring of parasite burden by measuring fluorescence signals. Therefore, this technique can be confidently used to directly monitor parasitic load and infection overtime and could be an excellent tool for in vitro and in vivo screening of anti-leishmanial drugs and vaccine efficiency. This is the first report of the use of the near-infrared fluorescence imaging technique for monitoring in vivo cutaneous leishmaniasis.
authors: Department of Virology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation (FIOCRUZ), Av. Professor Moraes Rego, s/n - Cidade Universitária, - 50.740-465 Recife, PE, Brazil. Electronic address: janacoliveira@hotmail.com.
authors: Janaina Correia
authors: JC
authors: Oliveira
authors: Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation (FIOCRUZ), Av. Professor Moraes Rego, s/n - Cidade Universitária, - 50.740-465 Recife, PE, Brazil. Electronic address: aline.caroline.bm@gmail.com.
authors: Aline Caroline
authors: AC
authors: da Silva
authors: Department of Virology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation (FIOCRUZ), Av. Professor Moraes Rego, s/n - Cidade Universitária, - 50.740-465 Recife, PE, Brazil. Electronic address: renatoasoliveira@hotmail.com.
authors: Renato Antonio Dos Santos
authors: RA
authors: Oliveira
authors: Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation (FIOCRUZ), Av. Professor Moraes Rego, s/n - Cidade Universitária, - 50.740-465 Recife, PE, Brazil. Electronic address: valeria@cpqam.fiocruz.br.
authors: Valéria Rêgo Alves
authors: VR
authors: Pereira
authors: Department of Virology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation (FIOCRUZ), Av. Professor Moraes Rego, s/n - Cidade Universitária, - 50.740-465 Recife, PE, Brazil. Electronic address: laura@cpqam.fiocruz.br.
authors: Laura Helena Vega Gonzales
authors: LH
authors: Gil
mesh_terms: D000818:Animals; D001483:Base Sequence; D016054:DNA, Protozoan; D004195:Disease Models, Animal; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D007259:Infrared Rays; D007894:Leishmania mexicana / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism* / Q000472:pathogenicity*; D016773:Leishmaniasis, Cutaneous / Q000175:diagnosis* / Q000469:parasitology; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics / Q000528:radiation effects; D051379:Mice; D008807:Mice, Inbred BALB C; D057054:Molecular Imaging / Q000379:methods; D061848:Optical Imaging / Q000379:methods*; D059208:Parasite Load; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000528:radiation effects*
chemical_list: D016054:DNA, Protozoan; D008164:Luminescent Proteins; D011994:Recombinant Proteins
keywords: BALB/c mouse; Cutaneous leishmaniasis; Leishmania amazonensis; Near-infrared molecular imaging; iRFP