Figure 7.
Figure 7. Intranasal vaccination, targeting LDCs, confers protection against CT-induced diarrhea. (A and B) CT-immunized mice (i.n., i.t., s.c., or p.o.) were challenged orally with CT 14 d after immunization. Cumulative data from at least three experiments is shown. Statistical comparisons to unimmunized (WT) mice are shown. (A) Quantification of the small intestinal weight. Horizontal bars, mean values; symbols, individual mice. (B) ELISA to quantify CT-specific IgA and IgG. (C) ELISPOT quantification of CT-specific IgA+ cells in the GI tract after i.n. and i.t. immunization with CT. Unimmunized mice are represented as WT. Cumulative data from two experiments, five to six mice pooled per experiment. Statistical comparisons to unimmunized (WT) mice are shown. (D) ELISA quantification of IgA in the stool and serum from mice immunized i.n., i.t., or s.l. Cumulative data from 10 mice is shown. Statistical comparisons between i.n./i.t. and s.l immunized mice are shown. (E and F) Mice immunized i.n. with 2 µg of CT, delivered either in a volume of 40 µl or 5 µl were compared. Cumulative data from at least three experiments is shown. (E) Quantification of the small intestinal weight. Horizontal bars, mean values; symbols, individual mice. Statistical comparisons to unimmunized (WT) mice are shown. (F) ELISA quantification of CT-specific IgA in stool and serum. Statistical comparisons to between the two volumes of i.n. vaccine are shown. (G) Flow cytometric quantification of IgA+ B cells in the lung and MedLN at steady state and after immunization with CT in WT or Batf3−/− (KO) mice. Cumulative data from two independent experiments is shown here with five mice per experiment. (H and I) CT-immunized WT and Batf3−/− (KO) mice were challenged orally with CT 14 d after immunization. (H) Quantification of the small intestinal weight after CT challenge. (I) ELISA to quantify CT-specific IgA in stool and serum. Horizontal bars, mean values; symbols, individual mice. Cumulative data from two independent experiments is shown. Statistical comparisons are made to immunized WT mice. (J) CT-immunized TCR−/− mice were challenged orally with CT 14 d after immunization. Small intestinal weight after CT challenge is demonstrated. (K and L) SPF or GF mice, immunized with CT were challenged orally after 14 d. Cumulative data from two experiments is shown. Statistical comparisons to SPF mice are shown. (K) Quantification of the small intestinal weight. Horizontal bars, mean values; symbols, individual mice. (L) ELISA quantification of CT-specific IgA in stool, lung, and serum. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Intranasal vaccination, targeting LDCs, confers protection against CT-induced diarrhea. (A and B) CT-immunized mice (i.n., i.t., s.c., or p.o.) were challenged orally with CT 14 d after immunization. Cumulative data from at least three experiments is shown. Statistical comparisons to unimmunized (WT) mice are shown. (A) Quantification of the small intestinal weight. Horizontal bars, mean values; symbols, individual mice. (B) ELISA to quantify CT-specific IgA and IgG. (C) ELISPOT quantification of CT-specific IgA+ cells in the GI tract after i.n. and i.t. immunization with CT. Unimmunized mice are represented as WT. Cumulative data from two experiments, five to six mice pooled per experiment. Statistical comparisons to unimmunized (WT) mice are shown. (D) ELISA quantification of IgA in the stool and serum from mice immunized i.n., i.t., or s.l. Cumulative data from 10 mice is shown. Statistical comparisons between i.n./i.t. and s.l immunized mice are shown. (E and F) Mice immunized i.n. with 2 µg of CT, delivered either in a volume of 40 µl or 5 µl were compared. Cumulative data from at least three experiments is shown. (E) Quantification of the small intestinal weight. Horizontal bars, mean values; symbols, individual mice. Statistical comparisons to unimmunized (WT) mice are shown. (F) ELISA quantification of CT-specific IgA in stool and serum. Statistical comparisons to between the two volumes of i.n. vaccine are shown. (G) Flow cytometric quantification of IgA+ B cells in the lung and MedLN at steady state and after immunization with CT in WT or Batf3−/− (KO) mice. Cumulative data from two independent experiments is shown here with five mice per experiment. (H and I) CT-immunized WT and Batf3−/− (KO) mice were challenged orally with CT 14 d after immunization. (H) Quantification of the small intestinal weight after CT challenge. (I) ELISA to quantify CT-specific IgA in stool and serum. Horizontal bars, mean values; symbols, individual mice. Cumulative data from two independent experiments is shown. Statistical comparisons are made to immunized WT mice. (J) CT-immunized TCR−/− mice were challenged orally with CT 14 d after immunization. Small intestinal weight after CT challenge is demonstrated. (K and L) SPF or GF mice, immunized with CT were challenged orally after 14 d. Cumulative data from two experiments is shown. Statistical comparisons to SPF mice are shown. (K) Quantification of the small intestinal weight. Horizontal bars, mean values; symbols, individual mice. (L) ELISA quantification of CT-specific IgA in stool, lung, and serum. *, P < 0.05; **, P < 0.01; ***, P < 0.001.

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