Increased survival and proliferation of the epidemic strain Mycobacterium abscessus subsp massiliense CRM0019 in alveolar epithelial cells

Autor(es)

Ribeiro, Giovanni Monteiro ; Matsumoto, Cristianne Kayoko ; Real, Fernando ; Teixeira, Daniela ; Duarte, Rafael Silva ; Mortara, Renato Arruda ; Leao, Sylvia Cardoso ; Carvalho-Wodarz, Cristiane de Souza

• Tipo de documento: Artigo Científico
• Fonte: BMC Microbiology; v. 17, SEP 13 2017.
• Citações Web of Science: 0

Resumo

Background: Outbreaks of infections caused by rapidly growing mycobacteria have been reported worldwide generally associated with medical procedures. Mycobacterium abscessus subsp. massiliense CRM0019 was obtained during an epidemic of postsurgical infections and was characterized by increased persistence in vivo. To better understand the successful survival strategies of this microorganism, we evaluated its infectivity and proliferation in macrophages (RAW and BMDM) and alveolar epithelial cells (A549). For that, we assessed the following parameters, for both M. abscessus CRM0019 as well as the reference strain M. abscessus ATCC 19977: internalization, intracellular survival for up 3 days, competence to subvert lysosome fusion and the intracellular survival after cell reinfection. Results: CRM0019 and ATCC 19977 strains showed the same internalization rate (approximately 30% after 6 h infection), in both A549 and RAW cells. However, colony forming units data showed that CRM0019 survived better in A549 cells than the ATCC 19977 strain. Phagosomal characteristics of CRM0019 showed the bacteria inside tight phagosomes in A549 cells, contrasting to the loosely phagosomal membrane in macrophages. This observation holds for the ATCC 19977 strain in both cell types. The competence to subvert lysosome fusion was assessed by acidification and acquisition of lysosomal protein. For M. abscessus strains the phagosomes were acidified in all cell lines; nevertheless, the acquisition of lysosomal protein was reduced by CRM0019 compared to the ATCC 19977 strain, in A549 cells. Conversely, in macrophages, both M. abscessus strains were located in mature phagosomes, however without bacterial death. Once recovered from macrophages M. abscessus could establish a new intracellular infection. Nevertheless, only CRM0019 showed a higher growth rate in A549, increasing nearly 10- fold after 48 and 72 h. Conclusion: M. abscessus CRM0019 creates a protective and replicative niche in alveolar epithelial cells mainly by avoiding phagosome maturation. Once recovered from infected macrophages, CRM0019 remains infective and displays greater intracellular growth in A549 cells compared to the ATCC 19977 strain. This evasion strategy in alveolar epithelial cells may contribute to the long survival of the CRM0019 strain in the host and thus to the inefficacy of in vivo treatment. (AU)

• Processo FAPESP: 09/14665-9 - O núcleo da célula hospedeira é necessário para a infecção por micobactérias?
• Beneficiário: Cristiane de Souza Carvalho
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 12/04913-8 - Estudo do comportamento intracelular de M. abscessus subsp. bolettii CRM0019
• Beneficiário: Giovanni Monteiro Ribeiro
• Modalidade de apoio: Bolsas no Brasil - Mestrado
• Processo FAPESP: 13/16018-6 - Sequenciamento e anotação do genoma da cepa epm 10906, uma nova espécie de micobactéria
• Beneficiário: Cristianne Kayoko Matsumoto
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado