Leishmania spp. parasite isolation through inoculation of patient biopsy macerates in interferon gamma knockout mice Leishmania spp.: isolamento de parasitos pela inoculação de macerados de biopsias de pacientes em camundongos deficientes em interferon gama

• Published in: Revista do Instituto de Medicina Tropical de São Paulo
• Main Authors: Milton Adriano Pelli de Oliveira, Alause da Silva Pires, Rosidete Pereira de Bastos, Glória Maria Collet de Araujo Lima, Sebastião Alves Pinto, Ledice Inácia de Araujo Pereira, Ana Joaquina Cohen Serique Pereira, Ises de Almeida Abrahamsohn, Miriam Leandro Dorta, Fátima Ribeiro-Dias
• Format: Article in Journal/Newspaper
• Language: English
• Published: Universidade de São Paulo (USP) 2010
• Subjects: Leishmania (Viannia) subgenus; Leishmania isolation; Interferon gamma deficient mice; Leishmaniasis; Leishmania identification; Experimental leishmaniasis; Arctic medicine. Tropical medicine; RC955-962; Infectious and parasitic diseases; RC109-216; Arctic
• Online Access: https://doi.org/10.1590/S0036-46652010000200004; https://doaj.org/article/65e2488fd6c1477d8308e41f94532045

Isolation of Leishmania parasite and species identification are important for confirmation and to help define the epidemiology of the leishmaniasis. Mice are often used to isolate pathogens, but the most common mouse strains are resistant to infection with parasites from the Leishmania (Viannia) subgenus. In this study we tested the inoculation of interferon gamma knockout (IFNγ KO) mice with biopsy macerates from Leishmania-infected patients to increase the possibility of isolating parasites. Biopsies from twenty five patients with clinical signs of leishmaniasis were taken and tested for the presence of parasites. Immunohistochemical assay (IHC) and conventional histopathology detected the parasite in 88% and 83% of the patients, respectively. Leishmania sp. were isolated in biopsy macerates from 52% of the patients by culture in Grace's insect medium, but 13% of isolates were lost due to contamination. Inoculation of macerates in IFNγ KO mice provides isolation of parasites in 31.8% of the biopsies. Most isolates belong to L. (Viannia) subgenus, as confirmed by PCR, except one that belongs to L. (Leishmania) subgenus. Our preliminary results support the use of IFNγ KO mice to improve the possibility to isolate New World Leishmania species. O isolamento e a identificação da espécie de parasito do gênero Leishmania são importantes para a confirmação e auxiliam na epidemiologia da leishmaniose. Os camundongos são freqüentemente utilizados para isolar patógenos, porém, as linhagens mais comuns de camundongos são resistentes à infecção por parasitos do subgênero Leishmania (Viannia). Neste estudo, avaliamos a inoculação de macerados de biópsias de pacientes infectados em camundongos deficientes do gene do interferon gama (IFNγ KO) como um método para aumentar a possibilidade de isolar Leishmania spp. Biópsias de 25 pacientes infectados com Leishmania sp. foram avaliadas para a presença de parasitos pelos métodos de imunohistoquímica (IHC) e histopatologia convencional. Os parasitos foram observados, ...