<i>A. vaginae</i> Induces Epithelial Expression of Soluble and Cell-Associated hBD2, a Protein That Attracts Primary Lymphocytes.

<p>A) Confluent monolayers of epithelial cells were inoculated with bacteria. Twenty-four hr post-inoculation, conditioned media were collected, clarified, and analyzed by ELISA to quantify concentrations of soluble hBD2 protein. Average MOI are: 5.8 for <i>L. johnsonii</i>, 2.5 for <i>G. vaginalis</i>, and 1.4 for <i>A. vaginae</i>. Results are averaged from 3 independent experiments, and asterisks indicate significant (p<0.001) increase over mock<i>-</i>treated condition. B) Confluent monolayers were inoculated with <i>A. vaginae</i>, or mock-inoculated. 24, 48, and 72 hr post-inoculation, cell monolayers were acid-extracted and analyzed by AU-PAGE western to quantify cell-associated hBD2 protein. A recombinant hBD2 protein standard (shown in ng per lane) was run alongside cell extracts for semi-quantitative comparison. Average MOI is 21. Shown is one example of three independent experiments. C) Unstimulated primary lymphocytes were isolated and evaluated for chemotaxis toward recombinant hBD2 protein. Chemotactic index is the ratio of migrated cells in hBD2-containing wells over vehicle-control wells, and significant increases over the matched vehicle condition are shown by three (p<0.001) asterisks.