Volume 118, Issue 4, Pages (April 2000)

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Presentation transcript:

Volume 118, Issue 4, Pages 749-759 (April 2000) Induction and maintenance of immune effector cells in the gastric tissue of mice orally immunized to Helicobacter pylori requires salivary glands Yasuhiko Shirai*,‡, Yoshio Wakatsuki*, Takashi Kusumoto‡, Mitsunori Nakata§, Masaru Yoshida*, Takashi Usui*, Tadahiko Iizuka‡, Toru Kita* Gastroenterology Volume 118, Issue 4, Pages 749-759 (April 2000) DOI: 10.1016/S0016-5085(00)70144-2 Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 1 Effect of sialoadenectomy on IgA secretion. One month after sialoadenectomy, the concentration of IgA in the (A) gastric juice, (B) gastric mucus, and (C) feces was measured. SMG+SLG, mice without sublingual and submandibular glands; SMG+SLG+PG, mice without sublingual, submandibular, and parotid glands. Each group consisted of 5 mice. Mean values + SD values are shown. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 8 Increase of (A) IgG1, (B) IgG2a, and (C) IgA antibodies specific to H. pylori in mice with sialoadenectomy after intragastric immunization with whole-cell sonicate of H. pylori and CT. SGE was prepared from mice intragastrically immunized with whole-cell sonicate of H. pylori and nonimmunized control mice. At each time of immunization in mice with sialoadenectomy, antigen preparation was mixed with SGE or PBS. Each experimental group consisted of 5 mice; mean optical density values + SD are shown. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 2 H. pylori–specific antibody responses after oral immunization. Mice were immunized by feeding sonicates of H. pylori and CT once a week for a total of 4 weeks. Concentrations of (A) secretory IgA and (B) serum IgG specific to H. pylori were measured at 2 weeks after the last immunization. O.I., oral immunization; SALX, sialoadenectomy. Mean values + SD are shown. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 3 Density of H. pylori colonizing the stomach at 1 month after inoculation. Mice were inoculated with a single dose of 5.7 × 107 of H. pylori suspended in 0.2 mL PBS by gastric intubation 2 weeks after the last immunization. Half of the stomach in each mouse was homogenized and processed for the culture of H. pylori. A positive culture was defined as visible colonies after 5 days of incubation. The colony-forming units were normalized by the weight of the stomach used for homogenate preparations. O.I., oral immunization; SALX, sialoadenectomy. Histograms indicate mean value + SD. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 4 H. pylori–specific antibody responses at 1 month after inoculation. H. pylori–specific (A) IgA and (B) IgM antibodies in gastric juice and (C) IgG antibodies in sera were measured as described in Materials and Methods. Histograms indicate mean value + SD. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 5 Density of H. pylori colonizing the stomach at 6 months after inoculation. Half of the stomach in each mouse was homogenized and processed for the culture of H. pylori. The colony-forming units were normalized by the weight of the stomach used for the homogenate preparations. Histograms indicate mean value + SD. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 6 The ratio of H. pylori–specific serum IgG1 to IgG2a determined (A) at the day of challenge and (B) 1 month after inoculation. Histograms indicate mean value + SD. *P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 7 Increase of (A and B) IgG1 and (C and D) IgG2a antibodies specific to H. pylori (A and C) 1 week before and (B and D) 1 month after challenge with H. pylori inoculation. Histograms indicate mean value + SD. P < 0.05. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions

Fig. 9 Grades of cell infiltration determined at 1 month after the challenge. Mice were killed at the indicated time points, and sections of the stomach were evaluated according to the criteria described in Materials and Methods by 2 pathologists blinded to the treatment of the mice. Mean score values ± SD are shown. ○, Group 1; □, group 2; ●, group 3; ■, group 4; ▴, group 5. Gastroenterology 2000 118, 749-759DOI: (10.1016/S0016-5085(00)70144-2) Copyright © 2000 American Gastroenterological Association Terms and Conditions