NIH Study Confirms Progressive
Clinical & Researched Lyme Position
- Lyme makes protective cysts-- "some spirochetes were arranged in cystic structures...
- Lyme changes its outer proteins to hide and survive in humans-- "invading spirochetes differentially expressed surface proteins...have been described for infected tissues in vivo.... to explore the gene activation responsible for the adaptation of B. burgdorferi to human tissue..."
It ain't what you don't know that gets you into trouble. It's what you know for sure that just ain't so. Mark Twain
Invasion of human tissue ex vivo by Borrelia burgdorferi
Borrelia burgdorferi sensu stricto is an etiological agent of Lyme disease. The lack of an adequate ex vivo system for human tissue infection is an obstacle to fully understanding the molecular mechanisms of invasion of tissue by B. burgdorferi and its adaptation within the human host. Here, we report on the development of such a system. We inoculated blocks of human tonsillar tissue with B. burgdorferi spirochetes, cultured them in a low-shear rotating wall vessel (RWV) bioreactor, and analyzed them using light and electron microscopy, nested polymerase chain reaction (PCR), and quantitative real-time PCR. Also, we evaluated the expression of the outer surface proteins (Osps) OspA and OspC by use of quantitative Western blotting. Light and electron microscopic analysis revealed multiple spirochetes localized extracellularly within the tissue, and their identity was confirmed by PCR. Quantification of spirochetes inside the RWV-cultured tonsillar tissue demonstrated that the number of B. burgdorferi exceeded the initial inoculum by an order of magnitude, indicating that spirochetes replicated in the tissue. Electron microscopic analysis showed that some spirochetes were arranged in cystic structures and that invading spirochetes differentially expressed surface proteins; both of these features have been described for infected tissues in vivo. The system we have developed can be used to study B. burgdorferi pathogenesis under controlled conditions ex vivo, in particular to explore the gene activation responsible for the adaptation of B. burgdorferi to human tissue that leads to Lyme disease.
Journal of Infectious Disease. 2005;15:1747-54. Epub 2005 Apr 6.
Duray PH, Yin SR, Ito Y, Bezrukov L, Cox C, Cho MS, Fitzgerald W, Dorward D, Zimmerberg J, Margolis L.
Department of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892, USA.