Abstract
Helicobacter pylori can transform from its normal helical bacillary morphology to a coccoid morphology. Since this coccoid form cannot be cultured in vitro, it has been speculated that it is a dormant form potentially involved in the transmission of H. pylori and in a patient's relapse after antibiotic therapy. In this study we determined the effects of aging, temperature, aerobiosis, starvation, and antibiotics on the morphologic conversion rate and culturability of H. pylori. Aerobiosis and the addition of a bactericidal antibiotic to the culture medium resulted in the highest conversion rate. During the conversion to coccoid forms, the cultures always lost culturability at the stage where 50% of the organisms were still in bacillary form; this result indicated that culturability and coccoid morphology are two separate but related entities. Independent of the conditions used to induce the conversion into coccoids, the morphological conversion was accompanied by several marked antigenic and ultrastructural changes. Also, both the total amounts and the integrity of RNA and DNA were significantly reduced in coccoid forms. With the potential-sensitive probe diOC(5)-3, a clear loss of membrane potential in coccoid forms was observed. Inhibition of protein or RNA synthesis by the addition of bacteriostatic antibiotics did not prevent the conversion to coccoid forms but resulted in an increased conversion rate. Hence, we conclude that conversion of H. pylori from the bacillary to the coccoid form is a passive process that does not require protein synthesis. Our data suggest that the coccoid form of H. pylori is the morphologic manifestation of bacterial cell death.
Full Text
The Full Text of this article is available as a PDF (907.5 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Benaissa M., Babin P., Quellard N., Pezennec L., Cenatiempo Y., Fauchère J. L. Changes in Helicobacter pylori ultrastructure and antigens during conversion from the bacillary to the coccoid form. Infect Immun. 1996 Jun;64(6):2331–2335. doi: 10.1128/iai.64.6.2331-2335.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Berry V., Jennings K., Woodnutt G. Bactericidal and morphological effects of amoxicillin on Helicobacter pylori. Antimicrob Agents Chemother. 1995 Aug;39(8):1859–1861. doi: 10.1128/aac.39.8.1859. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bode G., Malfertheiner P., Ströhle A., Mauch F., Nilius M., Ditschuneit H. Polymorphismus bei Helicobacter pylori--Schlüsselfunktion für Infektionsrezidive? Med Klin (Munich) 1992 Apr 15;87(4):179–184. [PubMed] [Google Scholar]
- Bode G., Mauch F., Malfertheiner P. The coccoid forms of Helicobacter pylori. Criteria for their viability. Epidemiol Infect. 1993 Dec;111(3):483–490. doi: 10.1017/s0950268800057216. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Boucher S. N., Slater E. R., Chamberlain A. H., Adams M. R. Production and viability of coccoid forms of Campylobacter jejuni. J Appl Bacteriol. 1994 Sep;77(3):303–307. doi: 10.1111/j.1365-2672.1994.tb03078.x. [DOI] [PubMed] [Google Scholar]
- Buck G. E. Campylobacter pylori and gastroduodenal disease. Clin Microbiol Rev. 1990 Jan;3(1):1–12. doi: 10.1128/cmr.3.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bukanov N. O., Berg D. E. Ordered cosmid library and high-resolution physical-genetic map of Helicobacter pylori strain NCTC11638. Mol Microbiol. 1994 Feb;11(3):509–523. doi: 10.1111/j.1365-2958.1994.tb00332.x. [DOI] [PubMed] [Google Scholar]
- Catrenich C. E., Makin K. M. Characterization of the morphologic conversion of Helicobacter pylori from bacillary to coccoid forms. Scand J Gastroenterol Suppl. 1991;181:58–64. [PubMed] [Google Scholar]
- Cellini L., Allocati N., Angelucci D., Iezzi T., Di Campli E., Marzio L., Dainelli B. Coccoid Helicobacter pylori not culturable in vitro reverts in mice. Microbiol Immunol. 1994;38(11):843–850. doi: 10.1111/j.1348-0421.1994.tb02136.x. [DOI] [PubMed] [Google Scholar]
- Cellini L., Allocati N., Di Campli E., Dainelli B. Helicobacter pylori: a fickle germ. Microbiol Immunol. 1994;38(1):25–30. doi: 10.1111/j.1348-0421.1994.tb01740.x. [DOI] [PubMed] [Google Scholar]
- Cellini L. Coccoid forms of Helicobacter pylori. J Infect Dis. 1996 May;173(5):1288–1289. doi: 10.1093/infdis/173.5.1288. [DOI] [PubMed] [Google Scholar]
- Chan W. Y., Hui P. K., Leung K. M., Chow J., Kwok F., Ng C. S. Coccoid forms of Helicobacter pylori in the human stomach. Am J Clin Pathol. 1994 Oct;102(4):503–507. doi: 10.1093/ajcp/102.4.503. [DOI] [PubMed] [Google Scholar]
- Cole S. P., Cirillo D., Kagnoff M. F., Guiney D. G., Eckmann L. Coccoid and spiral Helicobacter pylori differ in their abilities to adhere to gastric epithelial cells and induce interleukin-8 secretion. Infect Immun. 1997 Feb;65(2):843–846. doi: 10.1128/iai.65.2.843-846.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Eaton K. A., Catrenich C. E., Makin K. M., Krakowka S. Virulence of coccoid and bacillary forms of Helicobacter pylori in gnotobiotic piglets. J Infect Dis. 1995 Feb;171(2):459–462. doi: 10.1093/infdis/171.2.459. [DOI] [PubMed] [Google Scholar]
- Hazeleger W. C., Janse J. D., Koenraad P. M., Beumer R. R., Rombouts F. M., Abee T. Temperature-dependent membrane fatty acid and cell physiology changes in coccoid forms of Campylobacter jejuni. Appl Environ Microbiol. 1995 Jul;61(7):2713–2719. doi: 10.1128/aem.61.7.2713-2719.1995. [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]
- Moran A. P., Upton M. E. A comparative study of the rod and coccoid forms of Campylobacter jejuni ATCC 29428. J Appl Bacteriol. 1986 Feb;60(2):103–110. doi: 10.1111/j.1365-2672.1986.tb03366.x. [DOI] [PubMed] [Google Scholar]
- Moran A. P., Upton M. E. Factors affecting production of coccoid forms by Campylobacter jejuni on solid media during incubation. J Appl Bacteriol. 1987 Jun;62(6):527–537. doi: 10.1111/j.1365-2672.1987.tb02685.x. [DOI] [PubMed] [Google Scholar]
- NG L. K., Sherburne R., Taylor D. E., Stiles M. E. Morphological forms and viability of Campylobacter species studied by electron microscopy. J Bacteriol. 1985 Oct;164(1):338–343. doi: 10.1128/jb.164.1.338-343.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nilius M., Ströhle A., Bode G., Malfertheiner P. Coccoid like forms (CLF) of Helicobacter pylori. Enzyme activity and antigenicity. Zentralbl Bakteriol. 1993 Sep;280(1-2):259–272. doi: 10.1016/s0934-8840(11)80964-3. [DOI] [PubMed] [Google Scholar]
- Shahamat M., Mai U., Paszko-Kolva C., Kessel M., Colwell R. R. Use of autoradiography to assess viability of Helicobacter pylori in water. Appl Environ Microbiol. 1993 Apr;59(4):1231–1235. doi: 10.1128/aem.59.4.1231-1235.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Siegele D. A., Kolter R. Life after log. J Bacteriol. 1992 Jan;174(2):345–348. doi: 10.1128/jb.174.2.345-348.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Simoons-Smit I. M., Appelmelk B. J., Verboom T., Negrini R., Penner J. L., Aspinall G. O., Moran A. P., Fei S. F., Shi B. S., Rudnica W. Typing of Helicobacter pylori with monoclonal antibodies against Lewis antigens in lipopolysaccharide. J Clin Microbiol. 1996 Sep;34(9):2196–2200. doi: 10.1128/jcm.34.9.2196-2200.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sörberg M., Nilsson M., Hanberger H., Nilsson L. E. Morphologic conversion of Helicobacter pylori from bacillary to coccoid form. Eur J Clin Microbiol Infect Dis. 1996 Mar;15(3):216–219. doi: 10.1007/BF01591357. [DOI] [PubMed] [Google Scholar]
- Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet. 1983 Jun 4;1(8336):1273–1275. [PubMed] [Google Scholar]
- West A. P., Millar M. R., Tompkins D. S. Survival of Helicobacter pylori in water and saline. J Clin Pathol. 1990 Jul;43(7):609–609. doi: 10.1136/jcp.43.7.609-b. [DOI] [PMC free article] [PubMed] [Google Scholar]