首页> 美国卫生研究院文献>Clinical and Diagnostic Laboratory Immunology >The Campylobacter jejuni Dps Homologue Is Important for In Vitro Biofilm Formation and Cecal Colonization of Poultry and May Serve as a Protective Antigen for Vaccination
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The Campylobacter jejuni Dps Homologue Is Important for In Vitro Biofilm Formation and Cecal Colonization of Poultry and May Serve as a Protective Antigen for Vaccination

机译:空肠弯曲杆菌Dps同源物对于家禽的体外生物膜形成和盲肠定植很重要并且可以作为疫苗的保护性抗原

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摘要

In this work, we investigated the Campylobacter jejuni dps (DNA binding protein from starved cells) gene for a role in biofilm formation and cecal colonization in poultry. In vitro biofilm formation assays were conducted with stationary-phase cells in cell culture plates under microaerophilic conditions. These studies demonstrated a significant (>50%) reduction in biofilm formation by the C. jejuni dps mutant compared to that by the wild-type strain. Studies in poultry also demonstrated the importance of the dps gene in host colonization by C. jejuni. Real-time PCR analysis of mRNA extracted from the cecal contents of poultry infected with wild-type C. jejuni indicated that the dps gene is upregulated 20-fold during poultry colonization. Cecal colonization was greater than 5 log CFU lower in chicks infected with the dps mutant than chicks infected with the wild-type C. jejuni strain. Moreover, the dps mutant failed to colonize 75% of the chicks following challenge with 105 CFU. Preliminary studies were conducted in chicks by parenteral vaccination with a recombinant Dps protein or through oral vaccination with a recombinant attenuated Salmonella enterica strain synthesizing the C. jejuni Dps protein. No reduction in C. jejuni was noted in chicks vaccinated with the parenteral recombinant protein, whereas, a 2.5-log-unit reduction of C. jejuni was achieved in chicks vaccinated with the attenuated Salmonella vector after homologous challenge. Taken together, this work demonstrated the importance of Dps for biofilm formation and poultry colonization, and the study also provides a basis for continued work using the Dps protein as a vaccine antigen when delivered through a Salmonella vaccine vector.
机译:在这项工作中,我们调查了空肠弯曲杆菌dps(饥饿细胞的DNA结合蛋白)基因在禽类生物膜形成和盲肠定植中的作用。在微需氧条件下用细胞培养板中的固定相细胞进行体外生物膜形成测定。这些研究表明空肠弯曲菌dps突变体与野生型菌株相比,生物膜形成显着减少(> 50%)。家禽研究还证明了dps基因在空肠弯曲杆菌宿主定殖中的重要性。实时PCR分析从野生型空肠弯曲菌感染的家禽的盲肠中提取的mRNA表明,dps基因在家禽定殖过程中被上调了20倍。与感染野生型空肠弯曲杆菌菌株的雏鸡相比,感染dps突变体的雏鸡的盲肠定植降低了5 log CFU以上。此外,dps突变体在用10 5 CFU攻击后未能在75%的小鸡上定殖。通过用重组Dps蛋白进行肠胃外疫苗接种或通过合成空肠弯曲杆菌Dps蛋白的重组减毒沙门氏菌菌株口服疫苗对雏鸡进行了初步研究。接种肠胃外重组蛋白的雏鸡中空肠弯曲杆菌没有减少,而在同源攻击后接种减毒沙门氏菌载体的雏鸡中,空肠弯曲杆菌减少了2.5个对数单位。两者合计,这项工作证明了Dps在生物膜形成和家禽定殖中的重要性,并且该研究还为通过沙门氏菌疫苗载体递送Dps蛋白作为疫苗抗原的持续工作提供了基础。

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