Infect Immun 66(12): 5964-5971

Local Cytokine Response in <em>Helicobacter pylori</em>-Infected Subjects

Department of Medical Microbiology and Immunology, Göteborg University,^{and Department of Surgery, Sahlgrenska University Hospital,}^{Göteborg, Sweden}

^{Corresponding author. Mailing address: Department of Medical Microbiology and Immunology, Göteborg University, Guldhedsgatan 10A, S-413 46 Göteborg, Sweden. Phone: 46 31 342 62 16. Fax: 46 31 82 69 76. E-mail:}es.ug.oiborcim@mlohdnil.anirahtac.

Received 1998 Apr 24; Revisions requested 1998 Jul 15; Accepted 1998 Sep 17.

Abstract

The host immune response to Helicobacter pylori infection might be of importance with regard to the outcome of infection by this organism, e.g., to explain why only a proportion of infected subjects develop peptic ulcers. In this study we have analyzed the local response of different cytokines—i.e., the proinflammatory interleukin-1β, (IL-1β), IL-6, tumor necrosis factor alpha, and IL-8; the immunoregulatory gamma interferon (IFN-γ); and IL-4; and the anti-inflammatory transforming growth factor beta (TGF-β)—in antral biopsy specimens from H. pylori-infected duodenal ulcer (DU) patients and asymptomatic (AS) carriers (i.e., with chronic gastritis only). For comparison, biopsy specimens from uninfected healthy individuals were also analyzed. An immunohistochemical technique was used to allow quantification of the cytokine responses as well as identification of the cell types associated with the cytokine expression. We found that the levels of all of the studied cytokines except IL-4 were increased in the H. pylori-infected subjects compared to the levels in the healthy individuals. Our results indicate that the antral cytokine response is of the Th₁ type since IFN-γ, but not IL-4, was up-regulated both in H. pylori-infected DU patients and in AS carriers. However, there were no significant differences in either proinflammatory or immunoregulatory cytokine levels when H. pylori-infected subjects with and without peptic ulcers were compared. Some of the cytokines, particularly IL-1β and TGF-β, were also found in the gastric mucosae of healthy, uninfected subjects. We also showed that the gastric epithelium contributes substantially to the antral cytokine response of the proinflammatory cytokines IL-1β and IL-6 in addition to IL-8.

Abstract

Helicobacter pylori causes chronic antral gastritis and peptic ulcers and is associated with gastric adenocarcinoma and primary gastric lymphoma (30, 32, 34). H. pylori infection almost invariably causes chronic gastritis, but only a proportion of the infected subjects develop peptic ulcers. In addition to the possibility that some H. pylori strains might be more ulcerogenic than others, the nature of the host immune response might explain these different outcomes of infection by this organism. The local inflammation in H. pylori infection is characterized by infiltration of neutrophils and specific lymphocytes into the gastric mucosa as well as by increased production of several cytokines (10, 12, 16, 22, 43). Sometimes the infiltrating B and T cells form lymphoid follicles (15).

The immunoregulatory and proinflammatory cytokines induced by H. pylori may influence the nature of the local T-cell response. It is thought that helper T (Th) cells can be divided into two subsets, Th₁ and Th₂. The Th₁ subset promotes cell-mediated immunity by producing mainly interleukin-2 (IL-2) and gamma interferon (IFN-γ), and the Th₂ subset, which is important for antibody responses and also for down-regulation of chronic inflammatory reactions, produces IL-4, IL-5, IL-6, and IL-13 (37). The local Th cell response in H. pylori infection is generally held to be of the Th₁ type since the levels of IFN-γ, but not IL-4 and IL-5, have been shown to be increased in H. pylori-induced gastritis (20, 24). Furthermore, gastric H. pylori-specific T-cell clones isolated from peptic ulcer patients are more often of the Th₁ type than clones isolated from subjects with chronic gastritis only (12).

The gastric mucosal levels of the proinflammatory cytokines IL-1β, IL-6, IL-8, and tumor necrosis factor alpha (TNF-α) have been reported to be increased in H. pylori-infected subjects (26, 29, 43). In particular, the mucosal production of the neutrophil chemotactic and activating factor IL-8 has been suggested to play an important role in H. pylori-associated diseases (3, 8–10). H. pylori strains carrying the pathogenicity island, including the gene encoding the cytotoxin-associated protein (CagA), are more commonly isolated from duodenal ulcer (DU) patients than from infected subjects with chronic gastritis only, and infection with such strains has been found to be correlated with increased IL-8 production both in vivo and in vitro (10, 33). In addition, TNF-α and IL-1β can cause epithelial cell damage and induce epithelial IL-8 expression and therefore might also be of importance for the H. pylori-induced pathology (14). However, the inflammatory effects induced by the proinflammatory cytokines might be counteracted not only by IL-4 but also by locally produced transforming growth factor beta (TGF-β), which has been shown to have anti-inflammatory effects, e.g., inhibition of T cells, endothelial transmigration of neutrophils, and IL-8 production (11, 31, 39). The TGF-β response to H. pylori infection has not been extensively studied, however.

Most previous studies of H. pylori-induced cytokines have focused on detection of cytokine mRNA or quantification of protein in supernatants from in vitro cultures of gastric biopsy specimens, isolated gastric lymphocytes, or gastric epithelial cell lines (5, 19, 26, 29, 43). However, these approaches do not allow determination of either the localization or the nature of the cytokine-producing cells. In particular, the possible contribution of the antral epithelial cells to the cytokine response in H. pylori infection has hitherto not been extensively studied, except for that of IL-8 (8). In this study, we have therefore used immunohistochemical methods allowing identification as well as enumeration of the gastric cells associated with the expression of some of the cytokines which may be of importance for pathological mechanisms of H. pylori infection, i.e., IL-1β, IL-4, IL-6, IL-8, IFN-γ, TNF-α, and TGF-β. This has included a comparison of the expression of these cytokines in antral biopsy specimens from H. pylori-infected DU patients and subjects with chronic gastritis only (i.e., asymptomatic [AS] carriers), as well as from healthy, uninfected subjects, to evaluate whether there is any association between increased levels of one or more cytokines and peptic ulcer disease.

ACKNOWLEDGMENTS

Ulf and Jan Andersson are gratefully acknowledged for their kind and invaluable help in establishing the immunohistochemical technique in our laboratory. We thank Ingela Ahlstedt for excellent technical assistance and the staff at the Gastroenterological Endoscopy Unit, Sahlgrenska University Hospital, for skillful assistance with the biopsy samplings.

This study was financially supported by a grant from Astra Research Center, Boston, Mass.

ACKNOWLEDGMENTS

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