DNA fingerprints of Helicobacter pylori from mouth and antrum of patients with chronic ulcer dyspepsia

PBL (10’ in 200 AL) from patients and age-matched controls were cultured in presence of Staphylocoocus aureus CowanI (SAC) 0 0001%. IL-210 U/mL and IL-10 100 ng/mLwere added to some cultures. In parallel experiments, 10’ PBL were activated by CD40 system (XCD40, 10 x 10’ irradiated cells transfected with human Fc receptor [CDw32] plus CD40 monoclonal antibody 89 at 0.5 Ilgjml were used). Immunoglobulin in supernatants was measured on day 10 by isotype-specific enzyme-linked immunosorbent assay.

Secretion of IgG (ng/mL) patients with CVI

by peripheral

blood

lymphocytes from

SAC-activated B cells. Although SAC and high levels of IL-2 did not induce IgG production, IgG was produced when B cells were activated by SAC and IL-10. Failure of immunoglobulin production was not due to defective CD40 ligand-expression on T cells (data not shown). Our data show that IgG production in patients with CVI depends on the presence of IL-10. It is tempting to speculate that, in patients with CVI, B cells are functionally normal and that the failure of immunoglobulin production is due to an alteration in T-cell function, namely the impaired production of IL-10. S Zielen, Petra Bauscher, D Hofmann, S C Meuer Department of Paediatrics, J W Goethe-Universität, 6000 Frankfurt German Cancer Research Centre, Heidelberg

am

Main, FRG; and

1 Spickett GP, Farrant J. The role of lymphokines in common variable hypogammaglobulinaemia. Immunol Today 1989; 10: 192-94 2 Bryant A, Calver NC, Toubi E, Webster ADB, Farrant J. Classification of patients with CVI by B cell secretion of IgM and IgG in response to anti-IgM and interleukin-2. Clin Immunol Immunopathol 1990; 56: 239-48 3 Rousset F, Garcia E, Defrance T, et al. IL-10 is a potent growth and differentiation factor for activated human B lymphocytes. Proc Natl Acad Sci USA 1992; 89: 1890-93 4 Korthäuer U, Graf D, Mages HW, et al. Defective expression of T-cell CD40 ligand causes X-linked immune deficiency with hyper-IgM. Nature 1993; 361: 539-41 5 Banchereau J, Rousset F. Growing human B lymphocytes in the CD40 system. Nature 1991; 353: 678-79

Figure:

DNA

fingerprints of ribosomal RNA gene patterns

Hindlll restriction endonuclease digests of chromosomal DNA from H pylori. Fragments were identified by Southern blotting with a biotinylated cDNA derived from 16 + 23S ribosomal RNA of H pylori. Size marker (m) BstEll digest of bactenophage lambda DNA. Patients

1-5, p = plaque, and a = antral isolates

identity was confirmed by light and electron microscopy, and by urease activity. H pylori was obtained in 56 (89°,y) of gastric antral mucosal samples from ulcer subjects, and in 12 (19%) of these H pylori was also identified in cultures from dental plaque. H pylori was not cultured from the saliva of any subject. In 5 of the ulcer patients, identity of H pylori was confirmed with a specific cloned, random DNA probe.2 Paired strains from the mouth and antrum from each of these 5 patients were identical by DNA fingerprinting (ribotyping) (figure).2-4 Each patient was infected by a different strain of H pylori, except for patients 1 and 3, who had strains of similar DNA type. Failure to grow H pylori from saliva suggests that it is most unlikely that the endoscope carried H pylori from the antrum to the mouth. Furthermore the endoscope was separated from the teeth by a mouth guard throughout endoscopy. Dental plaque may be a reservoir for gastric reinfection by H pylori and sustained remission may depend on eradication of the organism from the mouth. Kashem

Khandaker, KR Palmer,

MA Eastwood

Gastrointestinal Unit, Department of Medicine, University of Edinburgh, Western General Hospital, Edinburgh EH4 2XU, UK

DNA fingerprints of Helicobacter pylori from mouth and antrum of patients with chronic ulcer dyspepsia SiR-Eradication of Helicobacter pylori from the stomach results in long-term remission of peptic ulcer, but eventually the stomach is recolonised and ulcer relapse follows. The source of reinfection is unknown, and we have explored the possibility that the oral cavity is an important reservoir for the

organism. Endoscopic gastric mucosal biopsy specimens, scrapings of dental plaque, and saliva were obtained from 51patients with active duodenal ulcers and 30 patients presenting with nonulcer dyspepsia. The presence of H pylori was sought by standard culture.’ Colonies resembling H pylori were subcultured

two to

four times

to

obtain pure cultures whose

AC Scott Central

Microbiological Laboratory, Western General Hospital

M Desai, RJ Owen National Collection of Type Cultures, Central Public Health

Laboratory,

London NW9

2

Skirrow MB. Campylobacter enteritis a "new" disease. BMJ 1977; 2: 9-11. Owen RJ, Hunton C, Bickley J, Moreno M, Lindon D. Ribosomal RNA gene restriction patterns of Helicobacter pylori: analysis and appraisal of HaeIII digests as a molecular typing system. Epidemiol

3

Desai

1

Infect 1992; 109:

4

35-47.

M, Linton D, Owen RJ, Cameron H, Stanley J. Genetic

diversity of Helicobacter pylori indexed with respect to clinical symptomatology, using a 16S rRNA and a species-specific DNA probe. J Applied Bacteriol (in press). Lindon D, Moreno M, Owen RJ, Stanley J. 16S rrn gene copy number in Helicobacter pylori and its application to molecular typing. J Applied Bacteriology 1992; 73: 501-06. 751