Jill - Bb tissue culture model

[Guest guest]

I think you mightve been looking for this one once to show me, or

maybe it was something else similar:

PMID: 15838803

Invasion of Human Tissue Ex Vivo by Borrelia burgdorferi

H. Duray,1 Shu-Rong Yin,2 Yoshinori Ito,2 Ludmila Bezrukov,2

Cheri ,2 Myong-Soon Cho,2 Fitzgerald,2 Dorward,3

Zimmerberg,2 and Leonid Margolis2

1Department of Pathology, National Cancer Institute, National

Institutes of Health, and 2National Aeronautics and Space

Administration/National Institutes of Health Center for Three-

Dimensional Tissue Culture, Laboratory of Cellular and Molecular

Biophysics, National Institute of Child Health and Human Development,

National Institutes of Health, Bethesda, land; 3National

Institute of Allergy and Infectious Diseases, National Institutes of

Health, Rocky Mountain Laboratories, Hamilton, Montana

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.