| Autor: |
de Paula NA; Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto14049-900, Brazil.; Dermatology Division, Department of Medical Clinics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil.; Reference Center for Sanitary Dermatology with Emphasis on Leprosy, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil., Leite MN; Dermatology Division, Department of Medical Clinics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil., de Faria Bertoluci DF; Department of Anatomic Pathology, Lauro de Souza Lima Institute, Bauru 17034-971, Brazil., Soares CT; Department of Anatomic Pathology, Lauro de Souza Lima Institute, Bauru 17034-971, Brazil., Rosa PS; Division of Research and Education, Lauro de Souza Lima Institute, Bauru 17034-971, Brazil., Frade MAC; Department of Cell and Molecular Biology and Pathogenic Bioagents, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto14049-900, Brazil.; Dermatology Division, Department of Medical Clinics, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil.; Reference Center for Sanitary Dermatology with Emphasis on Leprosy, Ribeirão Preto Medical School, University of São Paulo, Ribeirão Preto 14049-900, Brazil. |
| Abstrakt: |
The in vitro cultivation of M. leprae has not been possible since it was described as causing leprosy, and the limitation of animal models for clinical aspects makes studies on leprosy and bacteria-human host interaction a challenge. Our aim was to standardize the ex vivo skin model (hOSEC) to maintenance and study of M. leprae as an alternative animal model. Bacillary suspensions were inoculated into human skin explants and sustained in DMEM medium for 60 days. Explants were evaluated by RT-PCR-16SrRNA and cytokine gene expression. The viability and infectivity of bacilli recovered from explants (D28 and D60) were evaluated using the Shepard's model. All explants were RT-PCR-16SrRNA positive. The viability and infectivity of recovered bacilli from explants, analyzed after 5 months of inoculation in mice, showed an average positivity of 31%, with the highest positivity in the D28 groups (80%). Furthermore, our work showed different patterns in cytokine gene expression (TGF-β, IL-10, IL-8, and TNF-α) in the presence of alive or dead bacilli. Although changes can be made to improve future experiments, our results have demonstrated that it is possible to use the hOSEC to maintain M. leprae for 60 days, interacting with the host system, an important step in the development of experimental models for studies on the biology of the bacillus, its interactions, and drug susceptibility. |
