Drosophila melanogaster as an animal model for the study of Pseudomonas aeruginosa biofilm infections in vivo.
Drosophila melanogaster as an animal model for the study of Pseudomonas aeruginosa biofilm infections in vivo.
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DOI：
10.1371/journal.ppat.1002299
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发表时间：
2011-10
期刊：
影响因子：
6.7
通讯作者：
Lewenza S
中科院分区：
文献类型：
--
作者：
Mulcahy H;Sibley CD;Surette MG;Lewenza S
铜绿假单胞菌是一种机会致病菌，能够引起敏感宿主的急性和慢性感染。慢性绿脓杆菌感染被认为是由细菌生物膜引起的。生物膜是一种高度结构化的多细胞微生物群落，包裹在细胞外基质中，使其能够在宿主中长期存活。本研究的目的是开发一种动物模型，可以在果蝇宿主中进行铜绿假单胞菌生物膜感染的体内研究。在果蝇经口感染后24 h，铜绿假单胞菌生物膜定位于解剖的果蝇作物并在解剖的果蝇作物中可视化。这些生物膜具有特征性的聚集体结构和由DNA和胞外多糖组成的细胞外基质。从体内生长的生物膜中回收的铜绿假单胞菌细胞相对于体外生长的细胞具有增加的抗生素抗性。在体内，生物膜的形成依赖于pel胞外多糖基因的表达，因为pelB：：lux突变体不能形成生物膜。pelB：：lux突变体的毒力显著高于PAO 1，而超生物膜菌株（PAZHI 3）的毒力显著低于PAO 1，如感染后第14天受感染苍蝇的存活率所示。PAO 1和PAZHI 3菌株在农作物中的生物膜形成与感染后24 h诱导双蝶霉素、天蚕素A1和柔索霉素抗菌肽基因表达有关。相比之下，感染非生物膜形成菌株pelB：：lux导致果蝇中AMP基因表达降低。总之，这些结果为铜绿假单胞菌口腔感染果蝇期间宿主-病原体相互作用提供了新的见解，并突出了果蝇作为允许研究体内铜绿假单胞菌生物膜的感染模型的用途。 铜绿假单胞菌导致免疫系统受损的人严重感染。患有囊性纤维化的个体和医院患者特别容易受到铜绿假单胞菌感染。这种细菌对许多抗生素没有反应，使得这些感染难以治疗。铜绿假单胞菌可以作为自由漂浮的浮游细胞或称为生物膜的小菌落生长。铜绿假单胞菌形成生物膜的能力被认为有助于其引起慢性感染的能力。本研究的目的是开发一个简单的生物膜感染模型，使用果蝇（果蝇）。果蝇的免疫系统与脊椎动物的先天免疫系统相似。了解铜绿假单胞菌如何引起果蝇感染将有助于了解哺乳动物的毒力机制。在这项研究中，我们表明向果蝇喂食铜绿假单胞菌会导致生物膜感染，并且生物膜感染会诱导果蝇中抗菌肽免疫反应基因的表达。使用苍蝇生存作为衡量毒力，我们表明，生物膜感染的毒力比非生物膜感染。这些结果为铜绿假单胞菌感染期间宿主-病原体相互作用提供了新的见解。
Pseudomonas aeruginosa is an opportunistic pathogen capable of causing both acute and chronic infections in susceptible hosts. Chronic P. aeruginosa infections are thought to be caused by bacterial biofilms. Biofilms are highly structured, multicellular, microbial communities encased in an extracellular matrix that enable long-term survival in the host. The aim of this research was to develop an animal model that would allow an in vivo study of P. aeruginosa biofilm infections in a Drosophila melanogaster host. At 24 h post oral infection of Drosophila, P. aeruginosa biofilms localized to and were visualized in dissected Drosophila crops. These biofilms had a characteristic aggregate structure and an extracellular matrix composed of DNA and exopolysaccharide. P. aeruginosa cells recovered from in vivo grown biofilms had increased antibiotic resistance relative to planktonically grown cells. In vivo, biofilm formation was dependent on expression of the pel exopolysaccharide genes, as a pelB::lux mutant failed to form biofilms. The pelB::lux mutant was significantly more virulent than PAO1, while a hyperbiofilm strain (PAZHI3) demonstrated significantly less virulence than PAO1, as indicated by survival of infected flies at day 14 postinfection. Biofilm formation, by strains PAO1 and PAZHI3, in the crop was associated with induction of diptericin, cecropin A1 and drosomycin antimicrobial peptide gene expression 24 h postinfection. In contrast, infection with the non-biofilm forming strain pelB::lux resulted in decreased AMP gene expression in the fly. In summary, these results provide novel insights into host-pathogen interactions during P. aeruginosa oral infection of Drosophila and highlight the use of Drosophila as an infection model that permits the study of P. aeruginosa biofilms in vivo. Pseudomonas aeruginosa causes serious infections in people with compromised immune systems. Individuals with Cystic Fibrosis and hospital patients are particularly vulnerable to P. aeruginosa infections. This bacterium does not respond to many antibiotics, making these infections difficult to treat. P. aeruginosa can grow as free-floating planktonic cells or as microcolonies known as biofilms. The ability of P. aeruginosa to form biofilms is thought to contribute to their ability to cause chronic infections. The aim of this research was to develop a simple biofilm model of infection using the fruit fly (Drosophila melanogaster). The immune system of the fruit fly has similarities with the vertebrate innate immune system. Understanding how P. aeruginosa causes infections in Drosophila will aid in understanding virulence mechanisms in mammals. In this study we show that feeding P. aeruginosa to Drosophila results in a biofilm infection and biofilm infections induced expression of antimicrobial peptide immune response genes in the fly. Using fly survival as a measure of virulence we showed that biofilm infections were less virulent than non-biofilm infections. These results provide novel insight into host-pathogens interactions during P. aeruginosa infection.
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影响因子：
4.8
作者：
Hedengren-Olcott, M;Olcott, MC;Taylor, BJ
通讯作者：
Taylor, BJ
影响因子：
9.4
作者：
Ceri, H;Olson, ME;Buret, A
通讯作者：
Buret, A
DOI：
10.1111/j.1432-1033.1994.tb18730.x
发表时间：
1994-04-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
DIMARCQ, JL;HOFFMANN, D;HOFFMANN, JA
通讯作者：
HOFFMANN, JA
影响因子：
16
作者：
Hedengren, M;Åsling, B;Hultmark, D
通讯作者：
Hultmark, D
影响因子：
2.8
作者：
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通讯作者：
O'Gara, F