Tiansen LI, Chuangfu CHEN, Yueli WANG, Zhongchen MA, Huan ZHANG, Jihai YI, Yong WANG

Brucella abortus 2308’in Omp19 Geninin Silinmesi Faregil Makrofajında ve Farelerdeki Sağkalımını Azaltır

Bu çalışmanın amacı Brucella’nın dış zar proteini Omp19’un bakterinin patogenezindeki rolünü araştırmaktır. Bu araştırmada, Brucella 2308 ∆Omp19 delesyon mutantı oluşturularak faregil makrofajları RAW264.7 ve BALB/c farelerindeki hücre içi canlılığı değerlendirildi. Ayrıca faregil makrofajları RAW264.7’de Brucella’nın neden olduğu lizozomal füzyonu ve sitotoksisite de analiz edildi. Sonuçlar, Brucella 2308 ∆Omp19’un in vitro olarak immünoprotektif sitokin üretimini Brucella 2308’den daha fazla uyardığını ve RAW264.7 makrofajlarındaki hücre içi bakteri sayısının Brucella 2308 ∆Omp19 enfeksiyonunda Brucella 2308 enfeksiyonundan daha düşük olduğunu gösterdi. In vivo olarak, Brucella 2308 ∆Omp19, farelerde koruyucu bağışıklık tepkisine neden oldu ve bu delesyon mutantının dalak bakteri yükü, Brucella 2308’den daha düşüktü. Bunun yanı sıra Brucella 2308 ∆Omp19, makrofajlarda Brucella içeren vakuollerin ve lizozomların füzyonunu da uyardı. Brucella 2308 ∆Omp19, Brucella 2308 ile karşılaştırıldığında düşük virulansla konağın bağışıklık tepkisini aktive etmiştir ve zayıflatılmış canlı aşı için umut verici bir adaydır.

The Deletion of Omp19 Gene of Brucella abortus 2308 Reduces Its Survival in Mouse Macrophage and in Mice

The aim of this study is to investigate the role of outer membrane protein Omp19 of Brucella in its pathogenesis. In this research, the Brucella2308 ∆Omp19 deletion mutant was constructed and its intracellular survivability was evaluated in murine macrophages RAW264.7 and BALB/c mice. We also analyzed the lysosomal fusion induced by and cytotoxicity of Brucella in murine macrophages RAW264.7. The results showed that the Brucella 2308 ∆Omp19 induced higher levels of immunoprotective cytokines in vitro than Brucella 2308, and the number of intracellular bacteria in RAW264.7 macrophages was lower after Brucella 2308 ∆Omp19 infection than after Brucella 2308 infection. In vivo, Brucella 2308 ∆Omp19 induced protective immune response in mice, and the splenic bacterial load of this deletion mutant was lower than that of Brucella 2308. In addition, the Brucella 2308 ∆Omp19 also promoted the fusion of Brucella-containing vacuoles and lysosomes, in macrophages. Therefore, Brucella 2308 ∆Omp19 reduced virulence compared with Brucella 2308 and activate the immune response of the host, and is a promising candidate for a live attenuated vaccine.

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