THE EFFECT OF THE DELETION OF OPA 5 ON THE ABILITY OF N. GONORRHOEAE STRAIN MS11 TO TRANSCYTOSE A POLARIZED T84 EPITHELIAL CELL MONOLAYER
Stein, Daniel C
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<i>Neisseria gonorrhoeae<i> can express 11 different opacity proteins (Opa). Plasmids were constructed that allowed for the generation of strains containing deletions in each Opa-encoding-gene (amplified by PCR and cloned into pUC19). The Opa-encoding region was replaced with a spectinomycin resistance cassette and transformed into <i>N. gonorrhoeae<i>. The biological properties of one of these transformants (MS11 Opa5 delta s) was tested in a transcytosis assay. Wildtype MS11 was able to efficiently cross a polarized epithelial monolayer of T84 cells within 6 hours and occurred in both the apical to basolateral and the basolateral to apical directions. The number of cells that crossed the monolayer depended upon the dose of the inoculum. Transcytosis of MS11 Opa5 delta s occurred in the apical to basolateral direction, but was delayed. The data indicate that gonococci use multiple mechanisms of transcytosis, and the Opa 5 protein is important for rapid apical to basolateral transcytosis.