Abstract
Following exposure to Helicobacter pylori cells, epithelial cell lines secreted interleukin-6 (IL-6) and IL-8 but not tumor necrosis factor alpha. Purified IL-6 alone did not stimulate IL-8 production from the cell lines tested, indicating that IL-6 was not an intermediary in IL-8 induction. Enhanced IL-8 secretion occurred in a time- and dose-dependent manner. None of 12 antibiotics tested exhibited a significant effect on IL-8-inducing activity, suggesting that preformed antigens were responsible for stimulating IL-8 secretion in vitro. Live bacterial cells caused the highest level of stimulation. Proteinase-digested and heated (56 or 100 degrees C) cells had significantly reduced stimulatory activities. Purified H. pylori lipopolysaccharide, but not exopolysaccharide, stimulated low-level secretion of IL-8, but only at high concentrations, while a water-extracted H. pylori antigen preparation was strongly stimulatory for HEp-2 cells. No reduction in IL-8-stimulatory activity was observed for H. pylori mutants negative for urease activity, production of a major lipoprotein, and motility. The noncytotoxic strain CCUG 915 stimulated lower IL-8 levels than other isolates. However, the otherwise isogenic cytotoxin-negative mutant 17874 delta vacA (S. H. Phadnis, D. Ilver, L. Janzon, S. Normark, and T. U. Westblom, Infect. Immun. 62:1557-1565, 1994) had the same IL-8-stimulatory ability as the parent strain, suggesting that surface proteins other than the vacuolating cytotoxin are involved in IL-8 stimulation.
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- Agace W., Hedges S., Andersson U., Andersson J., Ceska M., Svanborg C. Selective cytokine production by epithelial cells following exposure to Escherichia coli. Infect Immun. 1993 Feb;61(2):602–609. doi: 10.1128/iai.61.2.602-609.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Austin J. W., Doig P., Stewart M., Trust T. J. Macromolecular structure and aggregation states of Helicobacter pylori urease. J Bacteriol. 1991 Sep;173(18):5663–5667. doi: 10.1128/jb.173.18.5663-5667.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Baggiolini M., Walz A., Kunkel S. L. Neutrophil-activating peptide-1/interleukin 8, a novel cytokine that activates neutrophils. J Clin Invest. 1989 Oct;84(4):1045–1049. doi: 10.1172/JCI114265. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Benabdelmoumene S., Dumont S., Petit C., Poindron P., Wachsmann D., Klein J. P. Activation of human monocytes by Streptococcus mutans serotype f polysaccharide: immunoglobulin G Fc receptor expression and tumor necrosis factor and interleukin-1 production. Infect Immun. 1991 Sep;59(9):3261–3266. doi: 10.1128/iai.59.9.3261-3266.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Blake M. S., Johnston K. H., Russell-Jones G. J., Gotschlich E. C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem. 1984 Jan;136(1):175–179. doi: 10.1016/0003-2697(84)90320-8. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- Colditz I., Zwahlen R., Dewald B., Baggiolini M. In vivo inflammatory activity of neutrophil-activating factor, a novel chemotactic peptide derived from human monocytes. Am J Pathol. 1989 Apr;134(4):755–760. [PMC free article] [PubMed] [Google Scholar]
- Correa P. Human gastric carcinogenesis: a multistep and multifactorial process--First American Cancer Society Award Lecture on Cancer Epidemiology and Prevention. Cancer Res. 1992 Dec 15;52(24):6735–6740. [PubMed] [Google Scholar]
- Cover T. L., Blaser M. J. Helicobacter pylori and gastroduodenal disease. Annu Rev Med. 1992;43:135–145. doi: 10.1146/annurev.me.43.020192.001031. [DOI] [PubMed] [Google Scholar]
- Cover T. L., Cao P., Lind C. D., Tham K. T., Blaser M. J. Correlation between vacuolating cytotoxin production by Helicobacter pylori isolates in vitro and in vivo. Infect Immun. 1993 Dec;61(12):5008–5012. doi: 10.1128/iai.61.12.5008-5012.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crabtree J. E., Peichl P., Wyatt J. I., Stachl U., Lindley I. J. Gastric interleukin-8 and IgA IL-8 autoantibodies in Helicobacter pylori infection. Scand J Immunol. 1993 Jan;37(1):65–70. doi: 10.1111/j.1365-3083.1993.tb01666.x. [DOI] [PubMed] [Google Scholar]
- Crabtree J. E., Taylor J. D., Wyatt J. I., Heatley R. V., Shallcross T. M., Tompkins D. S., Rathbone B. J. Mucosal IgA recognition of Helicobacter pylori 120 kDa protein, peptic ulceration, and gastric pathology. Lancet. 1991 Aug 10;338(8763):332–335. doi: 10.1016/0140-6736(91)90477-7. [DOI] [PubMed] [Google Scholar]
- Crabtree J. E., Wyatt J. I., Trejdosiewicz L. K., Peichl P., Nichols P. H., Ramsay N., Primrose J. N., Lindley I. J. Interleukin-8 expression in Helicobacter pylori infected, normal, and neoplastic gastroduodenal mucosa. J Clin Pathol. 1994 Jan;47(1):61–66. doi: 10.1136/jcp.47.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Darveau R. P., Hancock R. E. Procedure for isolation of bacterial lipopolysaccharides from both smooth and rough Pseudomonas aeruginosa and Salmonella typhimurium strains. J Bacteriol. 1983 Aug;155(2):831–838. doi: 10.1128/jb.155.2.831-838.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doig P., Austin J. W., Kostrzynska M., Trust T. J. Production of a conserved adhesin by the human gastroduodenal pathogen Helicobacter pylori. J Bacteriol. 1992 Apr;174(8):2539–2547. doi: 10.1128/jb.174.8.2539-2547.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Doig P., Trust T. J. Identification of surface-exposed outer membrane antigens of Helicobacter pylori. Infect Immun. 1994 Oct;62(10):4526–4533. doi: 10.1128/iai.62.10.4526-4533.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eckmann L., Kagnoff M. F., Fierer J. Epithelial cells secrete the chemokine interleukin-8 in response to bacterial entry. Infect Immun. 1993 Nov;61(11):4569–4574. doi: 10.1128/iai.61.11.4569-4574.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Huber A. R., Kunkel S. L., Todd R. F., 3rd, Weiss S. J. Regulation of transendothelial neutrophil migration by endogenous interleukin-8. Science. 1991 Oct 4;254(5028):99–102. doi: 10.1126/science.1718038. [DOI] [PubMed] [Google Scholar]
- Johnson J. A., Panigrahi P., Morris J. G., Jr Non-O1 Vibrio cholerae NRT36S produces a polysaccharide capsule that determines colony morphology, serum resistance, and virulence in mice. Infect Immun. 1992 Mar;60(3):864–869. doi: 10.1128/iai.60.3.864-869.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kadurugamuwa J. L., Clarke A. J., Beveridge T. J. Surface action of gentamicin on Pseudomonas aeruginosa. J Bacteriol. 1993 Sep;175(18):5798–5805. doi: 10.1128/jb.175.18.5798-5805.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kostrzynska M., O'Toole P. W., Taylor D. E., Trust T. J. Molecular characterization of a conserved 20-kilodalton membrane-associated lipoprotein antigen of Helicobacter pylori. J Bacteriol. 1994 Oct;176(19):5938–5948. doi: 10.1128/jb.176.19.5938-5948.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Leunk R. D., Johnson P. T., David B. C., Kraft W. G., Morgan D. R. Cytotoxic activity in broth-culture filtrates of Campylobacter pylori. J Med Microbiol. 1988 Jun;26(2):93–99. doi: 10.1099/00222615-26-2-93. [DOI] [PubMed] [Google Scholar]
- Linder H., Engberg I., Hoschützky H., Mattsby-Baltzer I., Svanborg C. Adhesion-dependent activation of mucosal interleukin-6 production. Infect Immun. 1991 Dec;59(12):4357–4362. doi: 10.1128/iai.59.12.4357-4362.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mai U. E., Perez-Perez G. I., Allen J. B., Wahl S. M., Blaser M. J., Smith P. D. Surface proteins from Helicobacter pylori exhibit chemotactic activity for human leukocytes and are present in gastric mucosa. J Exp Med. 1992 Feb 1;175(2):517–525. doi: 10.1084/jem.175.2.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mai U. E., Perez-Perez G. I., Wahl L. M., Wahl S. M., Blaser M. J., Smith P. D. Soluble surface proteins from Helicobacter pylori activate monocytes/macrophages by lipopolysaccharide-independent mechanism. J Clin Invest. 1991 Mar;87(3):894–900. doi: 10.1172/JCI115095. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Marshall B. J., Warren J. R. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet. 1984 Jun 16;1(8390):1311–1315. doi: 10.1016/s0140-6736(84)91816-6. [DOI] [PubMed] [Google Scholar]
- Matsushima K., Oppenheim J. J. Interleukin 8 and MCAF: novel inflammatory cytokines inducible by IL 1 and TNF. Cytokine. 1989 Nov;1(1):2–13. doi: 10.1016/1043-4666(89)91043-0. [DOI] [PubMed] [Google Scholar]
- Noach L. A., Bosma N. B., Jansen J., Hoek F. J., van Deventer S. J., Tytgat G. N. Mucosal tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-8 production in patients with Helicobacter pylori infection. Scand J Gastroenterol. 1994 May;29(5):425–429. doi: 10.3109/00365529409096833. [DOI] [PubMed] [Google Scholar]
- Nomura A., Stemmermann G. N., Chyou P. H., Kato I., Perez-Perez G. I., Blaser M. J. Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii. N Engl J Med. 1991 Oct 17;325(16):1132–1136. doi: 10.1056/NEJM199110173251604. [DOI] [PubMed] [Google Scholar]
- O'Toole P. W., Kostrzynska M., Trust T. J. Non-motile mutants of Helicobacter pylori and Helicobacter mustelae defective in flagellar hook production. Mol Microbiol. 1994 Nov;14(4):691–703. doi: 10.1111/j.1365-2958.1994.tb01307.x. [DOI] [PubMed] [Google Scholar]
- O'Toole P. W., Logan S. M., Kostrzynska M., Wadström T., Trust T. J. Isolation and biochemical and molecular analyses of a species-specific protein antigen from the gastric pathogen Helicobacter pylori. J Bacteriol. 1991 Jan;173(2):505–513. doi: 10.1128/jb.173.2.505-513.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Otterlei M., Sundan A., Skjåk-Braek G., Ryan L., Smidsrød O., Espevik T. Similar mechanisms of action of defined polysaccharides and lipopolysaccharides: characterization of binding and tumor necrosis factor alpha induction. Infect Immun. 1993 May;61(5):1917–1925. doi: 10.1128/iai.61.5.1917-1925.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parsonnet J., Friedman G. D., Vandersteen D. P., Chang Y., Vogelman J. H., Orentreich N., Sibley R. K. Helicobacter pylori infection and the risk of gastric carcinoma. N Engl J Med. 1991 Oct 17;325(16):1127–1131. doi: 10.1056/NEJM199110173251603. [DOI] [PubMed] [Google Scholar]
- Phadnis S. H., Ilver D., Janzon L., Normark S., Westblom T. U. Pathological significance and molecular characterization of the vacuolating toxin gene of Helicobacter pylori. Infect Immun. 1994 May;62(5):1557–1565. doi: 10.1128/iai.62.5.1557-1565.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Strieter R. M., Chensue S. W., Basha M. A., Standiford T. J., Lynch J. P., Baggiolini M., Kunkel S. L. Human alveolar macrophage gene expression of interleukin-8 by tumor necrosis factor-alpha, lipopolysaccharide, and interleukin-1 beta. Am J Respir Cell Mol Biol. 1990 Apr;2(4):321–326. doi: 10.1165/ajrcmb/2.4.321. [DOI] [PubMed] [Google Scholar]
- Sylvester I., Rankin J. A., Yoshimura T., Tanaka S., Leonard E. J. Secretion of neutrophil attractant/activation protein by lipopolysaccharide-stimulated lung macrophages determined by both enzyme-linked immunosorbent assay and N-terminal sequence analysis. Am Rev Respir Dis. 1990 Mar;141(3):683–688. doi: 10.1164/ajrccm/141.3.683. [DOI] [PubMed] [Google Scholar]
- Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tufano M. A., Rossano F., Catalanotti P., Liguori G., Capasso C., Ceccarelli M. T., Marinelli P. Immunobiological activities of Helicobacter pylori porins. Infect Immun. 1994 Apr;62(4):1392–1399. doi: 10.1128/iai.62.4.1392-1399.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tummuru M. K., Cover T. L., Blaser M. J. Cloning and expression of a high-molecular-mass major antigen of Helicobacter pylori: evidence of linkage to cytotoxin production. Infect Immun. 1993 May;61(5):1799–1809. doi: 10.1128/iai.61.5.1799-1809.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walz A., Peveri P., Aschauer H., Baggiolini M. Purification and amino acid sequencing of NAF, a novel neutrophil-activating factor produced by monocytes. Biochem Biophys Res Commun. 1987 Dec 16;149(2):755–761. doi: 10.1016/0006-291x(87)90432-3. [DOI] [PubMed] [Google Scholar]
- Yamamura M., Wang X. H., Ohmen J. D., Uyemura K., Rea T. H., Bloom B. R., Modlin R. L. Cytokine patterns of immunologically mediated tissue damage. J Immunol. 1992 Aug 15;149(4):1470–1475. [PubMed] [Google Scholar]