Scandinavian Journal of Gastroenterology

ISSN: 0036-5521 (Print) 1502-7708 (Online) Journal homepage: http://www.tandfonline.com/loi/igas20

Faecal Elastase Reflects Disease Activity in Active Ulcerative Colitis E. O. Adeyemi & H. J. F. Hodgson To cite this article: E. O. Adeyemi & H. J. F. Hodgson (1992) Faecal Elastase Reflects Disease Activity in Active Ulcerative Colitis, Scandinavian Journal of Gastroenterology, 27:2, 139-142, DOI: 10.3109/00365529209165434 To link to this article: http://dx.doi.org/10.3109/00365529209165434

Published online: 05 Aug 2009.

Submit your article to this journal

Article views: 14

View related articles

Citing articles: 4 View citing articles

Full Terms & Conditions of access and use can be found at http://www.tandfonline.com/action/journalInformation?journalCode=igas20 Download by: [University of Sydney Library]

Date: 25 March 2016, At: 22:06

Faecal Elastase Reflects Disease Activity in Active Ulcerative Colitis E. 0. ADEYEMI & H. J. F. HODGSON Gastroenterology Unit, Dept. of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, London, U.K.

Adeyemi EO, Hodgson HJF. Faecal elastase reflects disease activity in active ulcerative colitis. Scand

Downloaded by [University of Sydney Library] at 22:06 25 March 2016

. J Gastroenterol 1992, 27, 139-142

Alpha-1-proteinase inhibitor-bound elastase (EPIC) was measured in plasma and fresh stool samples from 20 patients with Crohn’s disease (CD), 16 patients with ulcerative colitis (UC), and 10 controls. Median EPIC values were significantly higher than normal in active CD and UC. EPIC was virtxally undetectable in the stool samples of control subjects. Median faecal EPIC in 14 patients with active CD (478 ng/ml) or 10 patients with active UC (1 159 ng/rnl) was significantly higher than in quiescent disease ( p < 0.05) and in control subjects ( p < 0.001 in each case). The difference in the median values between active CD and UC was not significant ( p = 0.065). The median faecal EPIC levels were identical in active UC (1159 ng/ml) and patients with large-bowel CD (LBCD) (1015 ng/ml) ( p = 0.9), and each was significantly higher than the value of 168 ng/ml in small-bowel CD (SBCD) ( p < 0.01 in each case). In active LBCD but not in SBCD, faecal EPIC correlated significantly with Crohn’s disease activity index ( R = 0.78, p < 0.05). plasma C-reactive protein (CRP) ( R = 0.9, p < 0.01), and erythrocyte sedimentation rate (ESR) ( R = 0.74, p < 0.05). In active UC, faecal EPIC correlated significantly with a numerical disease activity index ( R = 0.9, p < 0.01) but not with plasma EPIC and CRP, ESR, and leucocyte counts. Faecal EPIC values may be a better reflection of disease activity in active UC than plasma levels of markers of inflammation. Key words; Faecal elastase; inflammatory bowel disease Dr. E. 0. Adeyemi, Gastroenterology Unit, Dept. of Medicine, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, London, W12 ONN, U.K .

We have previously shown that levels of plasma polymorphonuclear leucocyte (PMNL) elastase, measured as elastase proteinase inhibitor complex (EPIC), are raised in patients with Crohn’s disease (CD) (1). EPIC levels are, however, either only modestly elevated or normal in active ulcerative colitis (UC), despite other objective and subjective indices of active disease. Similarly, for C-reactive protein (CRP) significant elevations occur in CD, but in UC only moderate elevation is observed in active disease (1,2). We surmised that measurement of EPIC in gut luminal contents, such as faeces, might be a better correlate of disease activity than plasma measurements, especially in cases of inflammatory bowel disease (IBD) which show a discordance between plasma EPIC levels and disease activity. This might be particularly relevant in UC, as plasma levels of EPIC are often only minimally elevated despite clinical and sigmoidoscopic evidence of disease activity. We have therefore measured stool EPIC and correlated its levels with Crohn’s disease activity index (CDAI) (3), other laboratory indices of inflammation, and a numerical disease activity index (NDAI). SUBJECTS, MATERIALS, AND METHODS

Fresh stool samples without urine admixture were obtained

from 10 control subjects (2 healthy volunteers, 6 patients with irritable bowel syndrome, and 2 patients with coeliac disease), 20 patients with CD (14 active and 6 inactive), and 16 patients with UC (10 active and 6 inactive; see Table 11). In the case of watery colitic stools, samples were drawn directly into large-bore 60-ml syringes and dispensed gently into pre-weighed universal Sterilin plastic containers, containing 100 1.11 of 100 mM ethylenediaminetetraacetic acid (EDTA) (EDTA was necessary to inhibit additional elastase secretion). Solid and semi-solid stool samples were taken with plastic spoons and placed in pre-weighed Sterilin plastic stool containers. The weight of each stool sample was noted. The volume (V,) of solid and semi-solid stool samples was derived by adding a known volume of normal saline (Vns),containing 100 mmol EDTA, and deducting this from the final volume (V,) thus: Vf = V, - Vns.The stool volume was needed to calculate the dilution factor. Watery colitic stools were centrifuged directly at 2000 g for 15min at 4°C. Each of the solid and semi-solid stool samples was suspended in 10 times its own volume of saIine/ EDTA, mixed gently with a whirl-mixer, and centrifuged at 20,OOOg for 30min at 4°C. The upper third of the stool supernatant was aspirated with a Pasteur pipette and used for enzyme-linked immunosorbent assay (ELISA) EPIC

140

E. 0 . Adeyemi & H. J . F. Hodgson

Table I. Items used in calculating the Crohn’s disease activity index (CDAI) in Crohn’s disease and a numerical disease activity index (NDAI) in ulcerative colitis Items 1) No. of liquid or very soft stools 2) Abdominal pain: sum of 7 daily ratings: 0 = no pain; 1 = mild; 2 = moderate; 3 = severe 3) General well-being: 0 = well; 1 = slightly below par; 2 = poor; 3 = very poor; 4 =

Downloaded by [University of Sydney Library] at 22:06 25 March 2016

terrible

4) Findings presumed related to Crohn’s disease a) Arthritis or arthralgia b) Skin or mouth lesions, such as pyoderma gangrenosum or erythema nodosum c) Iritis or uveitis d) Anal fissure, fistula, or perianal abscess e) Other bowel-related fistula, such as enterovesical f) Febrile episode exceeding 38°C during past week 5 ) Taking an antidiarrhoeal agent (1 = yes; 0 = no) 6) Abdominal mass (0 = absent; 2 = equivocal; 5 = present) 7 ) Haematocrit (M: 47 - Hct; F: 42 - Hct) 8) Body wt - 100 x (1 - (body wt/standard

Weighting factors 2 5

3 20

30 10

6

1

wt))

For the NDAI in ulcerative colitis, item 6 was substituted with bleeding per rectum.

measurement as described previously (4). In brief, the first antibody was a rabbit anti-elastase antibody, and the second a polyclonal antibody against human alpha-1-proteinase inhibitor (API) raised in the goat, and detection of EPIC was carried out with a third antibody, labelled with horseradish peroxidase (HRP). Stability of faecal E P I C at -20°C was assessed by determining the E P I C value in the same sample on the 3rd, 5th, and 7th day of storage. Plasma samples were also obtained from controls and patients for leucocyte (WBC) counts, EPIC, and C R P ELISA (4). Clinical assessment of disease activity in U C and CD and calculation of CDAI have been described previously (1). The NDAI for active UC, a slight modification of C D A I , derived by substituting bleeding per rectum for abdominal mass (Table I), was calculated on a weekly basis.

Statistics Mann-Whitney and Spearman’s correlation tests were used to process the different data groups in this study. RESULTS The standard curve used for the E P I C assay was linear from 1 to 50.0ng/ml, as described previously ( 5 ) . The lower detection limit was 1.0 ng/ml. The within-run variation was 8%, and the inter-assay coefficient of variation was 15% when EPIC measurements were carried out on a stool super-

natant in eight different assays. Recovery of added E P I C from liquid stool supernatant was greater than 95%. Median plasma E P I C and CRP levels of controls and IBD patients whose stool samples were studied are shown in Table 11. Each of the median plasma E P I C and C R P values in active C D was significantly higher than in active U C ( p = 0.003 for E P I C a n d p = 0.002 for CRP). Each of the median EPIC and C R P values was significantly higher than normal in active CD o r U C (Table 11). In the stool samples of normal controls, E P I C was hardly detectable, measuring only 20 ng/ml in one volunteer (median, 0 ng/ml; range, G 2 0 ng/ml). The median faecal EPIC levels in active CD and active U C patients were 478 and 1159 ng/ml, respectively. Each was significantly higher than normal ( p < 0.001), but the difference between the two median values did not reach the 0.05% confidence limit ( p = 0.065). When CD patients were divided into those having predominantly large-bowel (eight patients) and small-bowel (six patients) disease and compared with active UC patients, each of the median faecal E P I C values in active large-bowel C D (LBCD) (1015 ng/ml) and active U C (1159 ng/ml) was significantly higher than the median faecal E P I C in active small-bowel C D (SBCD) ( p < 0.001). The median faecal EPIC levels in active LBCD and UC were virtually identical (1015 ng/ml versus 1159 ng/ml; p = 0.9), and each of the median values, including that of active SBCD, was significantly higher than normal. Table 111 shows the median EPIC values in normal controls and active and inactive IBD. The median faecal E P I C level in active UC was significantly higher than in quiescent disease ( p < 0.01) with each being significantly higher than normal (Table Ill). Similarly, the median faecal E P I C value in active C D was significantly higher than in quiescent disease ( p = 0.03), and the median value in inactive CD was also significantly higher than normal ( p < 0.01) (Table 111). In active U C ( n = 10) faecal EPIC correlated significantly with a numerical disease activity index (NDAI) but not with

Table I1 Median. plasma elastase proteinase inhibitor complex (EPIC) and C-reactive protein (CRP) values in normal controls and patients with active and inactive Crohn’s disease (CD) and ulcerative colitis (UC), whose stool samples were studied Categories Controls (n = 10) Active CD ( n = 14) D* values Active UC ( n = 10) p*values Inactive CD ( n = 6 ) p* values Inactive UC ( n = 6) D* values

EPIC (ng/ml)

CRP ( m d )

75 (35-120) 1607 (92-291) 0.1 in each case 81 (56-115) >0.1 in each case

3.5$ (1-23)

p * comparison with normal controls;

30.5$ (1-131)

Faecal elastase reflects disease activity in active ulcerative colitis.

Alpha-1-proteinase inhibitor-bound elastase (EPIC) was measured in plasma and fresh stool samples from 20 patients with Crohn's disease (CD), 16 patie...
499KB Sizes 0 Downloads 0 Views