Carbonetti Nicholas H, Artamonova Galina V, Mays R Michael, Worthington Zoe E V
Department of Microbiology and Immunology, University of Maryland Medical School, Baltimore, Maryland 21201, USA.
Infect Immun. 2003 Nov;71(11):6358-66. doi: 10.1128/IAI.71.11.6358-6366.2003.
In this study, we sought to determine whether pertussis toxin (PT), an exotoxin virulence factor produced exclusively by Bordetella pertussis, is important for colonization of the respiratory tract by this pathogen by using a mouse intranasal infection model. By comparing a wild-type Tohama I strain to a mutant strain with an in-frame deletion of the ptx genes encoding PT (deltaPT), we found that the lack of PT confers a significant peak (day 7) colonization defect (1 to 2 log(10) units) over a range of bacterial inoculum doses and that this defect was apparent within 1 to 2 days postinoculation. In mixed-strain infection experiments, the deltaPT strain showed no competitive disadvantage versus the wild-type strain and colonized at higher levels than in the single-strain infection experiments. To test the hypothesis that soluble PT produced by the wild-type strain in mixed infections enhanced respiratory tract colonization by deltaPT, we coadministered purified PT with the deltaPT inoculum and found that colonization was increased to wild-type levels. This effect was not observed when PT was coadministered via a systemic route. Intranasal administration of purified PT up to 14 days prior to inoculation with deltaPT significantly increased bacterial colonization, but PT administration 1 day after bacterial inoculation did not enhance colonization versus a phosphate-buffered saline control. Analysis of bronchoalveolar lavage fluid samples from mice infected with either wild-type or deltaPT strains at early times after infection revealed that neutrophil influx to the lungs 48 h postinfection was significantly greater in response to deltaPT infection, implicating neutrophil chemotaxis as a possible target of PT activity promoting B. pertussis colonization of the respiratory tract.
在本研究中,我们试图通过使用小鼠鼻内感染模型来确定百日咳博德特氏菌独有的外毒素毒力因子百日咳毒素(PT)对于该病原体在呼吸道定殖是否重要。通过将野生型Tohama I菌株与编码PT的ptx基因发生框内缺失的突变菌株(deltaPT)进行比较,我们发现,在一系列细菌接种剂量范围内,缺乏PT会导致显著的峰值(第7天)定殖缺陷(1至2个对数(10)单位),且这种缺陷在接种后1至2天内就很明显。在混合菌株感染实验中,deltaPT菌株与野生型菌株相比没有竞争劣势,并且定殖水平高于单菌株感染实验。为了验证野生型菌株在混合感染中产生的可溶性PT增强了deltaPT在呼吸道定殖的假说,我们将纯化的PT与deltaPT接种物共同给药,发现定殖增加到了野生型水平。当通过全身途径共同给药PT时,未观察到这种效果。在接种deltaPT前长达14天经鼻内给予纯化的PT可显著增加细菌定殖,但在细菌接种后1天给予PT与磷酸盐缓冲盐水对照相比并未增强定殖。对感染野生型或deltaPT菌株的小鼠在感染后早期的支气管肺泡灌洗液样本分析显示,感染后48小时,肺部中性粒细胞对deltaPT感染的流入反应明显更大,这表明中性粒细胞趋化作用可能是PT促进百日咳博德特氏菌在呼吸道定殖的活性靶点。