Immunization of mice with urease vaccine affords protection against Helicobacter pylori infection in the absence of antibodies and is mediated by MHC class II-restricted responses - PubMed (original) (raw)

Comparative Study

Immunization of mice with urease vaccine affords protection against Helicobacter pylori infection in the absence of antibodies and is mediated by MHC class II-restricted responses

T H Ermak et al. J Exp Med. 1998.

Abstract

We examined the roles of cell- and antibody-mediated immunity in urease vaccine-induced protection against Helicobacter pylori infection. Normal and knockout mice deficient in major histocompatibility complex (MHC) class I, MHC class II, or B cell responses were mucosally immunized with urease plus Escherichia coli heat-labile enterotoxin (LT), or parenterally immunized with urease plus aluminum hydroxide or a glycolipid adjuvant, challenged with H. pylori strain X47-2AL, and H. pylori organisms and leukocyte infiltration in the gastric mucosa quantified. In an adjuvant/route study in normal mice, there was a direct correlation between the level of protection and the density of T cells recruited to the gastric mucosa. In knockout studies, oral immunization with urease plus LT protected MHC class I knockout mice [beta2-microglobulin (-/-)] but not MHC class II knockout mice [I-Ab (-/-)]. In B cell knockout mice [microMT (-/-)], vaccine-induced protection was equivalent to that observed in immunized wild-type (+/+) mice; no IgA+ cells were detected in the stomach, but levels of CD4(+) cells equivalent to those in the wild-type strain (+/+) were seen. These studies indicate that protection of mice against H. pylori infection by immunization with the urease antigen is dependent on MHC class II-restricted, cell-mediated mechanisms, and antibody responses to urease are not required for protection.

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Figures

Figure 1

Figure 1

Protection against H. pylori infection in Swiss-Webster mice immunized with recombinant urease using nonprotective, partially protective, and highly protective adjuvant and route combinations. (A) Gastric urease activity. (B) H. pylori CFUs from gastric mucosa. All groups were challenged with H. pylori. (Ctrl) Control mice were untreated. (IN/ day) IN urease daily over 14 d without adjuvant. (Al) Three biweekly s.c. immunizations with urease plus alum. (Bay) Three biweekly s.c. immunizations with urease plus Bay R 1005. (LT/Al) One IN immunization with urease plus LT followed by two s.c. immunizations with urease plus alum. (LT/Bay) One IN immunization with urease plus LT followed by two s.c. immunizations with urease plus Bay. (IN LT) Three biweekly IN immunizations with urease plus LT. Data points are from individual mice. In A, the bar represents the arithmetic mean of each group. In B, the bar represents the geometric mean CFU. Al and Bay groups showed significant protection after challenge as inferred from urease activity (P < 0.01), but at most a 10-fold decrease in bacterial density as determined by quantitative culture (Al, P = 0.05, and Bay, not significant, versus unimmunized controls; Al and Bay, P < 0.02, versus IN/day group). LT/Al and LT/Bay showed greater protection than Al or Bay alone (P < 0.04). IN LT was more protective than LT/Al (P < 0.03) or LT/Bay (P < 0.01). Wilcoxon/Kruskal-Wallis (rank sums) test.

Figure 2

Figure 2

Localization of CD4+ T cells in the gastric mucosa of urease-immunized, _H. pylori_–challenged mice. (a) Low-power field of gastric mucosa mounted on a slice of spleen (S). CD4+ cells localized at the junction region (J) of a mouse receiving s.c. urease. A, Antrum; C, corpus; and M, muscularis externa. (b) High-power field of CD4+ T cells in the gastric mucosa of a mouse receiving IN urease plus LT. T cells localized in the lamina propria between gastric pits and adjacent to the muscularis mucosa (arrowheads). Original magnifications: a, ×30; b, ×75.

Figure 3

Figure 3

T and B cell subsets in the gastric mucosa of H. pylori challenged mice as a function of adjuvant and route of administration. (A–C) Quantitative evaluation of CD4+, CD8+, and IgA+ cells. Mice immunized with urease plus adjuvant showed significant increases in T and B cells relative to unimmunized controls. IN/day group had no increase in T cells but did have increased IgA+ cells (P < 0.04). Some combination regimens were not significantly different from single adjuvant regimens. (D–F) Correlation of CD4+ cells, CD8+ cells, and IgA+ cells with geometric mean H. pylori CFUs.

Figure 4

Figure 4

Protection against H. pylori infection in wild-type (+/+), β2m (−/−), and I-Ab (−/−) knockout mice in response to challenge. (A) Gastric urease activity. (B) H. pylori CFUs from gastric mucosa. All groups were challenged with H. pylori. (Ctrl) Control mice received LT but no urease. (Al) Three biweekly s.c. immunizations with urease plus alum. (LT) Four weekly oral immunizations with urease plus LT. Data points are from individual mice. In A, the bar represents the arithmetic mean of each group. In B, the bar represents the geometric mean CFU. β2m (−/−) knockout mice receiving urease plus alum were not protected. Not shown, wild-type heterozygous (+/−) mice were protected after urease plus LT but not after urease plus alum. Lack of protection in these mice may reflect differences among (+/+), (+/−), and (−/−) strains.

Figure 5

Figure 5

Quantitative phenotypic analysis of the cellular infiltrate in the gastric mucosa of wild-type (+/+), β2m (−/−), and I-Ab (−/−) mice challenged with H. pylori. (A) CD4+ cells. (B) CD8+ cells. (C) CD103+ (αE-integrin+) cells. Most CD8+ and CD103+ cells in β2m (−/−) mice were gastric IEL. (D) IgA+ cells. All wild-type and β2m (−/−) groups had P < 0.05 versus controls, except the following alum groups: β2m CD4+ cells, wild-type and β2m IgA+ cells. I-Ab (−/−) mice showed no increase in T or B cells after immunization and challenge.

Figure 6

Figure 6

Protection against H. pylori infection in wild-type (+/+) and μMT (−/−) knockout mice. (A) Gastric urease activity. (B) H. pylori CFUs from gastric mucosa. All groups were challenged with H. pylori. (Ctrl) Mice were not immunized. (Al) Three biweekly s.c. immunizations with urease plus alum. (LT) Three biweekly oral immunizations with urease plus LT. Data points are from individual mice. In A, the bar represents the arithmetic mean of each group. In B, the bar represents the geometric mean CFU. LT groups, P < 0.01 versus unimmunized controls. In alum groups, only wild-type mice achieved a significant reduction in bacterial density (P < 0.02 versus unimmunized controls).

Figure 7

Figure 7

Quantitative evaluation of T cells, B cells, macrophages, and neutrophils in the gastric mucosa of wild-type (+/+) and μMT (−/−) mice challenged with H. pylori. (A) CD4+ cells. Wild-type, both groups P < 0.01 versus controls. μMT, both groups P = 0.05 versus controls. (B) CD8+ cells. Wild-type, both groups P < 0.01 versus controls. μMT mice, both groups not significant versus controls. (C) IgM+ cells (mostly small lymphocytes). (D) IgA+ cells (mostly plasma cells). (E) CD11b+ cells (macrophages). Wild-type, LT group, P < 0.03, alum group not significant versus controls. μMT, LT group not significant, alum group, P = 0.02 versus controls. (F) Ly-6G+ cells (neutrophils). Wild-type, both groups not significant versus controls. μMT mice, LT group, P < 0.05; alum group, P = 0.02 versus controls.

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