SPIRAL ORGANISMS IN THE BABOON STOMACH

634

prompted to assess possible differences between C pylori positive and negative gastric ulcers. 54 patients (mean age 59, range 36-78; 30 female) with untreated chronic benign gastric ulcer had at least two endoscopic biopsy specimens taken from uninvolved mucosa within 5 cm of the pylorus. The biopsy specimens were oriented on filter paper and immediately fixed in formol saline. Paraffin-processed sections were cut at three levels and stained by haematoxylin and eosin. A further section was taken from each set of specimens and stained by the undifferentiated Giemsa method ;’ these sections were examined "blind" by J. 1. W. for C pylori. The haematoxylin and eosin sections were examined "blind" by M. F. D. and specifically assessed for severity of reflux gastritis by a scoring system.2 Reflux gastritis has a characteristic histology comprising (1) foveolar hyperplasia, (2) oedema of the lamina propria, and (3) vasodilatation and congestion, and a paucity of both (4) acute and (5) chronic inflammatory cells. These five histological features were each graded 0 (normal or absent) to 3 (severe) and the sum of the grades was used as a composite reflux score for each patient. We have previously shown2,3 that a high reflux score (over 10) correlates closely with both hypochlorhydria (pH 4 or more) and increased total bile acid concentrations (over 1 mmol/1)-in gastric aspirates from fasting subjects. we were

evolution of gastric ulcer in C pylori positive patients may be related in some way to C pylori induced gastric mucosal injury, whereas in C pylori negative subjects duodenogastric reflux may be an important contributory factor. Gastroenterology Unit and University Departments of Pathology and Surgery, General Infirmary at Leeds,

H. J. O’CONNOR* M. F. DIXON J. I. WYATT A. T. R. AXON E. P. DEWAR

Leeds; Department of Pathology, St James’ University Hospital, Leeds, and Royal Naval Hospital, Haslar, Gosport *Present address

D. JOHNSTON

Queen Elizabeth Hospital, Edgbaston, Birmingham B15 2TH.

1. Rathbone

BJ, Wyatt JI, Heatley RV. Campylobacter pyloridis: a new factor in peptic ulcer disease? Gut 1986; 27: 635-41. 2. Dixon MF, O’Connor HJ, Axon ATR, King RFGJ, Johnston D. Reflux gastritis, a distinct histopathological entity? J Clin Pathol 1986; 39: 524-30. 3 O’Connor HJ, Dixon MF, Wyatt JI, et al. Effect of duodenal ulcer surgery and enterogastric reflux on Campylobacter pyloridis. Lancet 1986, ii 1178-81. 4. Rhodes J Etiology of gastric ulcer. Gastroenterology 1972, 63: 172-82.

SPIRAL ORGANISMS IN THE BABOON STOMACH

SIR,-We have found in the stomachs of baboons organisms to the new spiral bacterium from the human stomach reported by Miss Dent et al (July 11, p 96). This tightly spiralled similar

bacterium is morphologically distinct from true campylobacters that may occasionally be isolated from the stomach1,2 and from Campylobacter pylori.3 As yet this new organism has not been cultivated but we have isolated organisms that are structurally, biochemically, and serologically identical to Cpylori, from the same baboon stomachs (fig 1). From thin sections of the gastric mucosa, the newly recognised tightly spiralled organism has a gram-negative wall structure, is 4-8 l un long and 0-8 um wide, and has one complete tum of its spiral per 08 gm length. It has domed ends into both of which are inserted from two to twelve sheathed flagellar filaments. In some sections the outer membrane-like wall component is not closely adherent to the

Reflux scores in Cpylovi positive and negative gastric ulcer patients. 0

=

on

non-steroidal

anti-inflammatory

drugs. Horizontal bar

=

body spirals, producing a "cigar shaped" profile (fig 2). The organism lives in a mucus environment at the base of crypts and also seems to "bore" itself into parietal cells, where the invaginated cell membrane has a microvillous appearance (fig 3). This microvillous appearance has been observed by both Rollason et al4 and Chen et aP in the human stomach harbouring what were described as campylobacter-like organisms, although we have not observed such appearances in our examination of human biopsy specimens from which C pylori has been isolated. It is possible that this microvillous appearance is regularly associated only with this tightly spiralled organism. Classification of both this baboon organism and that recently found in man must await culture. In the pursuit of animal models of

median.

39 patients (72 %) (mean age 61, range 40-83; 21 female) were C pylori positive and 15 (mean age 56, range 36-78; 9 female) were negative. 1 patient in the negative group was taking oxytetracycline (to which C pylori is sensitive’) at the time of study and was therefore excluded from further analysis. Of the remaining 14 negative patients, 8 had a reflux score over 10 compared with only 5

of the C pylori positive patients (X2 = 10.9, p < 0001 ).

Furthermore,

C pylori negative patients had significantly higher reflux scores than positive patients (Wilcoxon rank sum test, p < 001; figure). 7 patients in the C pylori positive group (18%) compared with 2 (13%) in the negative group were taking non-steroidal antiinflammatory drugs at the time of study (p > 0-05). Thus the features that we believe constitute the histological picture of reflux gastritis were more severe in C pylori negative than in positive patients, and colonisation with the organism was uricommon in patients with a reflux score above 10. It is increasingly recognised that gastritis, from whatever cause, may predispose patients to peptic ulceration 4 We speculate that the

Fig 1= ‘C pylori like" organism from baboon stomach. Note

gently sinusoidal appearance (

x

13

000).

635

and its about equipotent as scavengers of .OH,3 which perhaps suggests that the specific action of 5-aminosalicylate in ulcerative colitis is unrelated to .OH scavenging. However, activated neutrophils also produce the powerful oxidant hypochlorous acid (HOCI), in a reaction dependent on the enzyme myeloperoxidase. 4 H OCI facilitates tissue damage at sites of inflammation by activating collagenase and inactivating o-antiprotease, thus permitting proteolytic enzymes to operate.4 Whereas sulphasalazine and sulphapyridine are poor scavengers of HOCI, 5-aminosalicylate, at concentrations present within the gut of patients on sulphasalazine therapy, is an effective HOCI scavenger.3 Other isomeric aminosalicylates also scavenge HOCI. Hence HOCI may be important in promoting tissue damage in ulcerative colitis, and scavengers of HOCI may be therapeutically useful.

damage

at

sites

of inflammation.2

Sulphasalazine

metabolites, sulphapyridine and 5-aminosalicylate,

Fig 2-Tightly spiralled bacteria from baboon stomach. Note the two forms of this organism: one with outer membrane-like component of the wall closely adherent to the spirals and one where this does not follow the spiral body, producing a cigar-shaped form ( x 13 000).

are

Department of Biochemistry, King’s College Strand Campus,

B. HALLIWELL

London WC2R 2LS 1. Babior BM. Oxidants from

phagocytes: Agents

of defence and destruction. Blood

1984; 64: 959-66. 2. Halliwell B, Gutteridge JMC. The importance of free radicals and catalytic metal ions in human disease. Molec Aspects Med 1985; 8: 89-193. 3. Aruoma OI, Wasil M, Halliwell B, Hoey BM, Butler J. The scavenging of oxidants by sulphasalazine and its metabolites: a potential contribution to their antiinflammatory effects? Biochem Pharmacol (in press). 4. Weiss SJ. Oxygen, ischaemia and inflammation. Acta Physiol Scand 1986; 548 (suppl): 9-37.

CLINICAL SIGNIFICANCE OF ANTI-IFN-&agr; ANTIBODY TITRES DURING INTERFERON THERAPY

Fig 3-Tightly spiralled bacteria in crypt lumen and in invagination of parietal cell. Note microvillous appearance of invaginated cell membrane (

x

5200).

C pylori infection and in the examination of human material, one be careful not to overlook or misidentify these morphologically distinct bacteria, which may coexist with C pylori, in both gram films of smeared mucus and in histological sections. must

We thank Dr N. Y. Haboubi for histological advice. Public Health Laboratory,

Withington Hospital, Manchester M20 8LR 1. 2 3.

4. 5

ALAN CURRY DENNIS M. JONES JOAN ELDRIDGE

N. Isolation from gastric epithelium of campylobacter-like bacteria that are distinct from "Campylobacter pyloridis". Lancet 1985; i. 111-12. Goodwm S, Blincow E, Armstrong J, McCulloch R, Collins D. Campylobacter pyloridis is unique. GCLO-2 is an ordinary campylobacter. Lancet 1985; ii: 38-39. Jones DM, Curry A, Fox AJ. Ultrastructure of a gastnc campylobacter-like organism "Campylobacter pyloridis". J Gen Microbiol 1985; 131: 2335-41 Rollason TP, Stone J, Rhodes JM Spiral organisms in endoscopic biopsies of the human stomach J Clin Pathol 1984; 37: 23-26. Chen XG, Correa P, Offerhaus J, et al Ultrastructure of the gastnc mucosa campylobacter-like organisms. AJCP 1986; 86: 575-82.

Kasper G, Dickgiesser

harboring

SULPHASALAZINE AND OXIDANT SCAVENGING IN ULCERATIVE COLITIS

SIR,-Your editorial (June 6, p 1299) discusses the possibility that sulphasalazine acts as a radical scavenger. Activated neutrophils, which are abundant at sites of inflammation, generate the superoxide radical (0) and hydrogen peroxide (HZOZ).1 O2 and HZOz can interact, in the presence of suitable transition metal catalysts, to form the highly reactive hydroxyl radical, .OH, which is thought to contribute to tissue

SiR,—In 1981 Vallbracht et aP reported a patient who had neutralising antibodies to natural interferon-beta (IFN-(3) during IFN-(3 therapy. Since then, anti-IFN antibodies have been detected in several clinical trials, which mainly used recombinant IFN-oc2a or IFN-oc2b. The proportion of patients who had antibodies during treatment varied from 0 to 44%, depending on the disease, dose schedule, and IFN preparation.2-5 It has been claimed4 that IFN-a2a leads to a significantly higher number of antibody positive patients than IFN-a2b. In a study in which patients with chronic myelogenous leukaemia were treated with 5 x 106 IU IFN-a2b subcutaneously thrice weekly, we have monitored anti-IFN-a antibodies before and during therapy by three different assays. Sera were tested in an ELISA with peroxidase-labelled IFN-CXzb,6 in an immunoradiometric assay (IRMA) similar to that described by Protzman et al,7 and in a neutralisation bioassay with Wish cells and vesicular stomatitis virus.8 Detectable amounts of anti-IFN-ot antibodies developed in 5 of 25 treated patients. After emergence of the IFN antibodies, these 5 patients did not respond to or relapsed during IFN therapy. The leucocyte and thrombocyte counts in their peripheral blood decreased over 1 to 9 months after the start of therapy but then increased. In contrast, the leucocyte and blast counts of the 20 antibody negative patients continuously decreased over at least 12 months (figure). The data demonstrate the clinical significance of IFN antibodies, which appear to neutralise IFN at least in part and thus interfere with IFN treatment. In 4 of the anti-IFN-ot positive patients measurable changes in the antibody titre were observed during treatment. The specific titre began to fall rapidly 1 h after IFN injection and reached its pre-injection level slowly after 48 h. After cessation of treatment, a continuous rise of the specific antibodies was observed for up to 5 days. We conclude that the emergence of anti-IFN antibodies affects the clinical response to IFN, that anti-IFN titres are underestimated when determined in patients still under treatment, and that the time at which blood is collected is critical for detecting anti-IFN antibodies. We therefore suggest that every patient treated with IFN should be monitored for anti-IFN antibodies. Blood specimens for anti-IFN assay should be drawn either