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Enhanced M1 macrophage polarization in human helicobacter pylori-associated atrophic gastritis and in vaccinated mice.

Quiding-Järbrink M, Raghavan S, Sundquist M - PLoS ONE (2010)

Bottom Line: Gene expression analysis of stomach tissue and sorted macrophages revealed that gastric macrophages were polarized to M1 after H. pylori infection, and this process was substantially accelerated by prior vaccination.Human H. pylori infection was characterized by a mixed M1/M2 polarization of macrophages.However, in H. pylori-associated atrophic gastritis, the expression of inducible nitric oxide synthase was markedly increased compared to uncomplicated gastritis, indicative of an enhanced M1 macrophage polarization in this pre-malignant lesion.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.

ABSTRACT

Background: Infection with Helicobacter pylori triggers a chronic gastric inflammation that can progress to atrophy and gastric adenocarcinoma. Polarization of macrophages is a characteristic of both cancer and infection, and may promote progression or resolution of disease. However, the role of macrophages and their polarization during H. pylori infection has not been well defined.

Methodology/principal findings: By using a mouse model of infection and gastric biopsies from 29 individuals, we have analyzed macrophage recruitment and polarization during H. pylori infection by flow cytometry and real-time PCR. We found a sequential recruitment of neutrophils, eosinophils and macrophages to the gastric mucosa of infected mice. Gene expression analysis of stomach tissue and sorted macrophages revealed that gastric macrophages were polarized to M1 after H. pylori infection, and this process was substantially accelerated by prior vaccination. Human H. pylori infection was characterized by a mixed M1/M2 polarization of macrophages. However, in H. pylori-associated atrophic gastritis, the expression of inducible nitric oxide synthase was markedly increased compared to uncomplicated gastritis, indicative of an enhanced M1 macrophage polarization in this pre-malignant lesion.

Conclusions/significance: These results show that vaccination of mice against H. pylori amplifies M1 polarization of gastric macrophages, and that a similar enhanced M1 polarization is present in human H. pylori-induced atrophic gastritis.

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Identification of gastric lamina propria DCs.(A) Flow cytometric analyses of CD11c+MHC-II+ cells in the gastric lamina propria of individual mice revealed two F4/80− DC populations with differential expression of CD103 (lower quadrants in the F4/80 versus CD103 dot plot). (B) Expression of CD103 and CD11c by gastric lamina propria cells. The gate for CD103+ DCs is indicated. Isotype, isotype-matched control antibody for CD103. (C) Expression of MHC-II, CD8α, F4/80 and CD11b by gated CD11c+CD103+ cells. Dotted lines show staining with isotype controls. (D) Expression of CD11b by CD103− and CD103+ DCs. DCs were gated as described in A. (E) Frequency of CD103+ DCs in the gastric lamina propria at the indicated time points after H. pylori infection. Data represents means ± SEM of 7–15 individual mice. LP, lamina propria.
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pone-0015018-g002: Identification of gastric lamina propria DCs.(A) Flow cytometric analyses of CD11c+MHC-II+ cells in the gastric lamina propria of individual mice revealed two F4/80− DC populations with differential expression of CD103 (lower quadrants in the F4/80 versus CD103 dot plot). (B) Expression of CD103 and CD11c by gastric lamina propria cells. The gate for CD103+ DCs is indicated. Isotype, isotype-matched control antibody for CD103. (C) Expression of MHC-II, CD8α, F4/80 and CD11b by gated CD11c+CD103+ cells. Dotted lines show staining with isotype controls. (D) Expression of CD11b by CD103− and CD103+ DCs. DCs were gated as described in A. (E) Frequency of CD103+ DCs in the gastric lamina propria at the indicated time points after H. pylori infection. Data represents means ± SEM of 7–15 individual mice. LP, lamina propria.

Mentions: To characterize gastric DCs, we first identified a population of CD11c+MHC-II+ cells (Fig. 2A). When these cells were further analyzed for expression of F4/80 and the αE integrin chain CD103, half of the CD11c+MHC-II+ cells were identified as F4/80+ macrophages (Fig. 2A). However, among the CD11c+MHC-II+ cells that lacked expression of F4/80, two populations of putative DCs with differential expression of CD103 could be distinguished (Fig. 2A). Due to the many fluorochromes required we chose to only characterize the gastric CD103+ DCs further, since these cells have been implicated as important antigen presenting cells in mucosal tissues [18]. Gastric CD103+ DCs were easily identified by staining for CD11c and CD103 (Fig. 2B). The gastric CD103+ DCs expressed high levels of MHC-II, and consisted of a CD11blow and a CD11bhigh subset (Fig. 2C). In comparison, the gastric CD103− DCs were all CD11bhigh (Fig. 2D). In addition, the CD103+ DCs lacked expression of CD8α and F4/80 (Fig. 2C). However, the frequency of CD103+ DCs did not change significantly in the gastric mucosa after infection (Fig. 2E).


Enhanced M1 macrophage polarization in human helicobacter pylori-associated atrophic gastritis and in vaccinated mice.

Quiding-Järbrink M, Raghavan S, Sundquist M - PLoS ONE (2010)

Identification of gastric lamina propria DCs.(A) Flow cytometric analyses of CD11c+MHC-II+ cells in the gastric lamina propria of individual mice revealed two F4/80− DC populations with differential expression of CD103 (lower quadrants in the F4/80 versus CD103 dot plot). (B) Expression of CD103 and CD11c by gastric lamina propria cells. The gate for CD103+ DCs is indicated. Isotype, isotype-matched control antibody for CD103. (C) Expression of MHC-II, CD8α, F4/80 and CD11b by gated CD11c+CD103+ cells. Dotted lines show staining with isotype controls. (D) Expression of CD11b by CD103− and CD103+ DCs. DCs were gated as described in A. (E) Frequency of CD103+ DCs in the gastric lamina propria at the indicated time points after H. pylori infection. Data represents means ± SEM of 7–15 individual mice. LP, lamina propria.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2990716&req=5

pone-0015018-g002: Identification of gastric lamina propria DCs.(A) Flow cytometric analyses of CD11c+MHC-II+ cells in the gastric lamina propria of individual mice revealed two F4/80− DC populations with differential expression of CD103 (lower quadrants in the F4/80 versus CD103 dot plot). (B) Expression of CD103 and CD11c by gastric lamina propria cells. The gate for CD103+ DCs is indicated. Isotype, isotype-matched control antibody for CD103. (C) Expression of MHC-II, CD8α, F4/80 and CD11b by gated CD11c+CD103+ cells. Dotted lines show staining with isotype controls. (D) Expression of CD11b by CD103− and CD103+ DCs. DCs were gated as described in A. (E) Frequency of CD103+ DCs in the gastric lamina propria at the indicated time points after H. pylori infection. Data represents means ± SEM of 7–15 individual mice. LP, lamina propria.
Mentions: To characterize gastric DCs, we first identified a population of CD11c+MHC-II+ cells (Fig. 2A). When these cells were further analyzed for expression of F4/80 and the αE integrin chain CD103, half of the CD11c+MHC-II+ cells were identified as F4/80+ macrophages (Fig. 2A). However, among the CD11c+MHC-II+ cells that lacked expression of F4/80, two populations of putative DCs with differential expression of CD103 could be distinguished (Fig. 2A). Due to the many fluorochromes required we chose to only characterize the gastric CD103+ DCs further, since these cells have been implicated as important antigen presenting cells in mucosal tissues [18]. Gastric CD103+ DCs were easily identified by staining for CD11c and CD103 (Fig. 2B). The gastric CD103+ DCs expressed high levels of MHC-II, and consisted of a CD11blow and a CD11bhigh subset (Fig. 2C). In comparison, the gastric CD103− DCs were all CD11bhigh (Fig. 2D). In addition, the CD103+ DCs lacked expression of CD8α and F4/80 (Fig. 2C). However, the frequency of CD103+ DCs did not change significantly in the gastric mucosa after infection (Fig. 2E).

Bottom Line: Gene expression analysis of stomach tissue and sorted macrophages revealed that gastric macrophages were polarized to M1 after H. pylori infection, and this process was substantially accelerated by prior vaccination.Human H. pylori infection was characterized by a mixed M1/M2 polarization of macrophages.However, in H. pylori-associated atrophic gastritis, the expression of inducible nitric oxide synthase was markedly increased compared to uncomplicated gastritis, indicative of an enhanced M1 macrophage polarization in this pre-malignant lesion.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology and Immunology, University of Gothenburg, Gothenburg, Sweden.

ABSTRACT

Background: Infection with Helicobacter pylori triggers a chronic gastric inflammation that can progress to atrophy and gastric adenocarcinoma. Polarization of macrophages is a characteristic of both cancer and infection, and may promote progression or resolution of disease. However, the role of macrophages and their polarization during H. pylori infection has not been well defined.

Methodology/principal findings: By using a mouse model of infection and gastric biopsies from 29 individuals, we have analyzed macrophage recruitment and polarization during H. pylori infection by flow cytometry and real-time PCR. We found a sequential recruitment of neutrophils, eosinophils and macrophages to the gastric mucosa of infected mice. Gene expression analysis of stomach tissue and sorted macrophages revealed that gastric macrophages were polarized to M1 after H. pylori infection, and this process was substantially accelerated by prior vaccination. Human H. pylori infection was characterized by a mixed M1/M2 polarization of macrophages. However, in H. pylori-associated atrophic gastritis, the expression of inducible nitric oxide synthase was markedly increased compared to uncomplicated gastritis, indicative of an enhanced M1 macrophage polarization in this pre-malignant lesion.

Conclusions/significance: These results show that vaccination of mice against H. pylori amplifies M1 polarization of gastric macrophages, and that a similar enhanced M1 polarization is present in human H. pylori-induced atrophic gastritis.

Show MeSH
Related in: MedlinePlus