Methods for Rapid Transfer and Localization of Lyme Disease Pathogens Within the Tick Gut
Methods for Rapid Transfer and Localization of Lyme Disease Pathogens Within the Tick Gut
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DOI：
10.3791/2544
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发表时间：
2011-02-01
影响因子：
1.2
通讯作者：
Pal, Utpal
中科院分区：
文献类型：
--
作者：
Kariu, Toru;Coleman, Adam S.;Pal, Utpal
莱姆病是由螺旋体病原体伯氏疏螺旋体感染引起的，其在自然界中通过蜱-啮齿动物感染循环维持（1）。已经开发了蜱传小鼠模型2以在实验室中研究莱姆病。而未感染蜱可感染B。通过用感染的老鼠喂养它们，蜕皮过程需要几个星期到几个月才能完成。因此，开发更快速有效的蜱感染技术，如基于显微注射的程序，是研究莱姆病的重要工具（3，4）。该程序只需要几个小时就能产生受感染的蜱虫，并允许控制蜱虫群中等量螺旋体的传递。这对于生成B尤其重要。通过使用小鼠的自然喂养过程感染伯氏菌的蜱不能确保100%的感染率，并且可能导致喂养的蜱中病原体负荷的变化。此外，可以使用显微注射来感染B的蜱。在减毒株不能在小鼠中建立感染并且因此不能被蜱自然获得的情况下，分离出伯氏螺旋体（5）。该技术也可用于将各种其他生物材料递送到蜱中，例如特异性抗体或双链RNA（6）。在这篇文章中，我们将演示用体外生长的B显微注射若虫蜱。burgdorferi。我们还将描述一种使用共聚焦免疫荧光显微镜在蜱肠道中定位莱姆病病原体的方法。
Lyme disease is caused by infection with the spirochete pathogen Borrelia burgdorferi, which is maintained in nature by a tick-rodent infection cycle(1). A tick-borne murine model 2 has been developed to study Lyme disease in the laboratory. While naive ticks can be infected with B. burgdorferi by feeding them on infected mice, the molting process takes several weeks to months to complete. Therefore, development of more rapid and efficient tick infection techniques, such as a microinjection-based procedure, is an important tool for the study of Lyme disease(3,4). The procedure requires only hours to generate infected ticks and allows control over the delivery of equal quantities of spirochetes in a cohort of ticks. This is particularly important as the generation of B. burgdorferi infected ticks by the natural feeding process using mice fails to ensure 100% infection rate and potentially results in variation of pathogen burden amongst fed ticks. Furthermore, microinjection can be used to infect ticks with B. burgdorferi isolates in cases where an attenuated strain is unable to establish infection in mice and thus can not be naturally acquired by ticks(5). This technique can also be used to deliver a variety of other biological materials into ticks, for example, specific antibodies or double stranded RNA(6). In this article, we will demonstrate the microinjection of nymphal ticks with in vitro-grown B. burgdorferi. We will also describe a method for localization of Lyme disease pathogens in the tick gut using confocal immunofluorescence microscopy.