Article Figures & Data
Figures
- FIG. 1.
Schematic representation of SpaP-DTA fusion proteins. The sizes of SpaP (open bar) and DTA (solid bar) are indicated by the number of amino acids (a.a.). Hatched bars indicate the C-terminal cell wall-anchoring domain of SpaP.
- FIG. 2.
Expression of recombinant DTA by S. gordonii. (A) Recombinant DTA proteins on the cell surface (solid bars) and in culture supernatants (open bars) were detected by ELISA. A405 readings were determined from diluted samples equivalent to culture optical densities at 600 nm of 0.25. (B) Recombinant DTA proteins were detected in cell extracts and culture supernatants by Western immunoblotting. Lane 1, S. gordonii DTA; lane 2, S. gordonii DTA2; lane 3, S. gordonii DTA3; lane 4, cell extracts of S. gordonii DL-1 (negative control). (C) Proteins were extracted from cells by previously described methods (13). S, proteins from culture supernatant fluids. Prestained protein size markers are shown on the left. In both panels, the proteins were detected with a mouse anti-DT antibody.
- FIG. 3.
Western immunoblots showing recognition of native DT by antiserum obtained from mice immunized with heat-killed recombinant S. gordonii. DT (1 μg) was separated on an SDS-10% PAGE gel. Mouse antisera: lane 1, S. gordonii DTA; lane 2, S. gordonii DTA2; lane 3, S. gordonii DTA3. P, preimmune serum; I, immune serum; +, anti-DT antiserum; M, prestained protein size markers (sizes shown in kilodaltons).
- FIG. 4.
Immune responses to S. gordonii DTA2 in BALB/c mice. (A) Anti-DT and anti-CTB antibodies elicited by intranasal immunization with live S. gordonii DTA2 in the presence of CTB or with CTB alone. (B) Anti-DT antibodies induced by oral colonization with S. gordonii DTA2. The values shown are mean titers ± standard error for individual animal samples. In the CTB-alone and SL3 groups, the absence of a bar indicates that an immune response was not detected. VW, vaginal wash.
