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Rheumatology 2001; 40: 15-23
© 2001 British Society for Rheumatology

Correlation between the immune responses to collagens type I, III, IV and V and Klebsiella pneumoniae in patients with Crohn's disease and ankylosing spondylitis

H. Tiwana1, R. S. Natt12, R. Benitez-Brito1, S. Shah1, C. Wilson1, S. Bridger2, M. Harbord3, M. Sarner3 and A. Ebringer1,4

1 Division of Life Sciences, Infection and Immunity Group, King's College London, Stamford Street, London
2 Department of Medicine, King's College Hospital, London
3 Department of Medicine, Middlesex Hospital, UCL School of Medicine, London
4 Department of Rheumatology, Middlesex Hospital, UCL School of Medicine, London, UK


   Abstract
Top
 Abstract
Introduction
 Patients and methods
 Results
 Discussion
 References
 
Background. Increased levels of collagen types I, III and V are found in strictures of patients with Crohn's disease (CD) compared with normal gut tissue. Type IV collagen is present in the basement membranes, basal lamina, retina and cornea. Elevated levels of antibody to Klebsiella pneumoniae are found in both active CD and active ankylosing spondylitis (AS) patients compared with healthy controls.

Methods. Reactivities for immunoglobulin class-specific antibodies (IgM, IgG and IgA) against collagen types I, III, IV, V and whole K. pneumoniae were measured by ELISA in nine patients with early CD and 10 with late CD from King's College Hospital and 12 late CD patients and 36 HLA-B27-positive AS patients from Middlesex Hospital and was compared with values for 26 healthy controls from the Blood Transfusion Service in London.

Results. Levels of class-specific IgM, IgG and IgA antibodies to collagen types I, III, IV, V and K. pneumoniae were significantly elevated in early and late CD patients compared with healthy controls (P<0.001). Levels of IgM, IgG antibody to the four collagen types and K. pneumoniae were also significantly elevated (P<0.001) in AS patients compared with healthy controls. In addition, the level of IgA antibody to K. pneumoniae was elevated in AS patients (P<0.001). Furthermore, a positive correlation between antibody levels to collagen types I, III, IV and K. pneumoniae was demonstrated in both early and late CD patients and in those with AS, whilst a positive correlation to type V was found in early CD.

Conclusion. The role of K. pneumoniae and anti-collagen antibodies in the aetiopathogenesis of CD and AS requires further study.

KEY WORDS: Crohn's disease, Ankylosing spondylitis, Collagens I, III, IV and V, Klebsiella pneumoniae.


   Introduction
Top
 Abstract
 Introduction
Patients and methods
 Results
 Discussion
 References
 
Crohn's disease (CD) is an inflammatory disorder affecting the gastrointestinal tract, particularly the ileocaecal region. It is associated with increased collagen deposition, which leads to the development of strictures, and these often occur early in the course of the disease [1]. The pathogenesis of stricture formation in the gastrointestinal tract is unknown. The narrowing of the lumen of the bowel due to the accumulation of collagen can cause partial or complete obstruction, which then requires surgical correction. Immunohistochemical studies have shown that the collagens involved in CD are usually of types I, III and V [2], which are present in the lamina propria, muscularis mucosae and muscularis propria respectively [3].

Ankylosing spondylitis (AS) can be considered a reactive arthritis following Klebsiella pneumoniae infection in HLA-B27-positive patients, on the basis of findings of raised anti-Klebsiella antibodies and the presence of molecular mimicry between the HLA-B27 molecule and bacterial antigens [4,5]. Amino acid homology has been described between the extracellular starch-induced enzyme pullulanase of K. pneumoniae and collagen types I, III and IV [6]. Histological changes in thigh muscle biopsies of AS patients have been reported [7] in which type III collagen is present. Type IV collagen is found in the basement membranes, basal lamina, retina and cornea [8]. Furthermore, elevated levels of IgG and IgA antibodies to both type I and type IV collagen have been demonstrated in AS patients [6]. Crohn's disease-like lesions have also been shown to be present in AS patients [9]. Spondyloarthropathy occurs in at least 20% of inflammatory bowel disease (IBD) patients [10]. Specific elevations of antibodies to Klebsiella in CD patients were first reported in 1987 [11]. This observation was subsequently confirmed when an extensive study involving three pathogenic microbes and 10 anaerobic isolates of the normal bowel flora showed elevated titres of antibodies to Klebsiella in both CD and AS patients, but no such elevations were found against Proteus, Escherichia coli or any of 10 anaerobic bacteria [12]. Previously, our group has shown that RA patients have elevated levels of antibodies to Proteus mirabilis and not to Klebsiella, whereas AS patients have elevated immune responses to Klebsiella and not to P. mirabilis [13]. Furthermore, class-specific antibodies to capsular serotypes of K. pneumoniae have been reported to be increased in both active CD patients and active AS patients [14].

The aims of this study were three-fold: (i) to determine whether the levels of antibodies to collagen types I, III, IV and V and to K. pneumoniae are increased in active patients with CD and AS compared with healthy controls; (ii) to determine if the pattern of antibody class responses to these collagen types and K. pneumoniae is different in patients with late CD compared with those with an early diagnosis of the disease; and (iii) most importantly, to determine if there is a correlation between anti-collagen and K. pneumoniae antibodies among CD and AS patients.


   Patients and methods
Top
 Abstract
 Introduction
 Patients and methods
Results
 Discussion
 References
 
Patients
In total, serum samples from 93 subjects were studied. Sera were obtained from nine patients with early-stage CD (five males, four females) (mean age 30 yr, range 15–77 yr); 10 patients with late-stage CD (seven males, three females) (mean age 31 yr, range 20–52 yr). The early- and late-stage CD patients attended the Gastroenterology Outpatients Clinic at King's College Hospital. Sera were also obtained from 12 patients with late CD (eight males, four females) (mean age 46 yr, range 17–79 yr) attending the Gastroenterology Outpatients Clinic at the Middlesex Hospital. Sera from 36 unrelated HLA-B27-positive patients with active AS (24 males, 12 females) (mean age 46 yr, range 31–70 yr) satisfying the Rome–New York diagnostic criteria and having an erythrocyte sedimentation rate (ESR) >15 mm/h were obtained from the AS Research Clinic at the Middlesex Hospital, London. All AS patients were being treated with sulphasalazine-EN (0.5–2.0 g daily) and a low-starch diet [15]. In addition, sera from 26 control subjects were supplied by the Blood Transfusion Service in London (14 males, 12 females) (mean age 41 yr, range 30–59 yr). The general characteristics of the study groups are shown in Table 1Go.


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  		TABLE 1. General characteristics of the study groups

 

Diagnosis of Crohn's disease
The diagnosis of CD was made using the standard St Marks criteria [16], with a Harvey–Bradshaw score >=3 [17], and a biochemical/permeability test was performed on each patient. The early-stage group (duration of disease <2 yr) consisted of patients newly diagnosed with CD after January 1997. Late-stage diagnosis of CD was defined by the physicians’ assessment of patients with a medical history of CD and a date of diagnosis before January 1997.

CRP and ESR determinations
Both C-reactive protein (CRP) and ESR levels were measured in the CD patients. At the time the blood sample was obtained, six of nine early CD patients had a CRP level >10 mg/l compared with 15 of 22 patients with late CD diagnosis. Five of nine early CD patients and 18 of 22 with late CD had an ESR level >15 mm/h.

Preparation of collagen
Collagen types I, III, IV and V (Sigma, Poole, UK) were dissolved in NaHCO3 (0.1 M) containing NaCl (0.5 M) to a final collagen concentration of 1 mg/ml.

Preparation of K. pneumoniae
A K. pneumoniae isolate of capsular type K50 (NTCC-9170) was kindly provided by Dr T. Pitt (Division of Hospital Infection, Central Public Health Laboratory, Colindale). The method of preparation of the bacterial suspension is described in detail elsewhere [13]. Briefly, the cultures were grown aerobically in nutrient broth, harvested and suspended in 0.15 M phosphate-buffered saline (PBS), pH 7.4. A stock solution was prepared to give an optical density (OD) reading at 540 nm on the Cecil 500 Double Beam Spectrophotometer that gave a suspension of 6x108 bacteria per ml [13].

Enzyme-linked immunosorbent assay (ELISA)
ELISA studies on collagen types I, III, IV and V.
The ELISA protocol used in the study was similar to that used by several investigators in this field of study in that antibody levels are reported in OD units rather than concentrations [5,13]. Ninety-six-well, flat-bottomed polystyrene microtitre plates (Dynatech, Billingshurst, UK) were coated with 200 µl of 1 mg/ml collagen solution [collagen+carbonate buffer (0.05 M), pH 9.6] and incubated overnight at 4°C. The plates were then washed three times with PBS–Tween and 300 µl of blocking solution in PBS was added to each well [2% (w/v) casein, 0.1% (v/v) Tween 20 in PBS], and the plates were incubated for 2 h at room temperature. Excess blocking solution was removed and 200-µl serum samples (test or control) diluted 1/200 in PBS–Tween were added in duplicate. Plates were incubated at room temperature for 2 h, followed by the addition of 200 µl peroxidase-conjugated rabbit anti-human IgM, IgG or IgA serum (Dako) diluted 1/500 in PBS–Tween, and incubated for a further 2 h at room temperature. Development of the colorimetric assay also took place at room temperature for 20 min after the addition of 200 µl per well of 0.5 mg/ml 2,2'-azinobis (3-ethylbenz-thiazoline-6-sulphonic acid (Sigma) in citrate phosphate buffer, pH 4.1, containing 0.98 mM H2O2 (Sigma). The reaction was stopped with 100 µl of 2 mg/ml sodium fluoride (Sigma) and the OD was measured at a wavelength of 630 nm with a micro-ELISA plate reader (Dynatech MR600). All assays were carried out under code, in that the tester did not know whether the sera came from patients or controls, and the results were expressed as OD units±S.E.

Bacterial ELISA.
K. pneumoniae suspended in carbonate buffer (0.05 M), pH 9.6, was adsorbed onto 96-well flat bottomed microtitre plates overnight at 4°C in 200-µl volumes. The plates were then washed and saturated with a blocking solution [0.1% (w/v) bovine serum albumin, 0.05% (v/v) Tween 20] in PBS and incubated for 1 h at 37°C. The rest of the ELISA procedure was carried out as mentioned above except that the plates were incubated at 37°C for 1 h after the addition of the individual serum samples and the peroxidase-conjugated antibodies.

Statistical analysis.
The mean OD units for IgM, IgG and IgA antibodies against K. pneumoniae and collagen types I, III, IV and V in the various patient groups and the control group were compared using Student's t-test. Differences in levels of the immuno-globulin classes were assessed by comparing the number of individuals in each group having OD units greater than the 95% confidence limits for the control group (one-tailed test), using the {chi}2 test with Yates’ correction. The correlation coefficient (r) was calculated with the analytical program Minitab, release 5.1 (Pearson's correlation coefficient), and the reproducibility of the antibody assay for each sample was measured by calculating the coefficient of variation. Individual samples were plotted (scatter graph) and histograms drawn using the statistical package Prism 3.0.


   Results
Top
 Abstract
 Introduction
 Patients and methods
 Results
Discussion
 References
 
Levels of IgM, IgG and IgA antibodies to K. pneumoniae (K50) and to each of the four collagen types were measured by ELISA in CD and AS patients and healthy controls. Data for the CD patients from the two centres were combined and analysed as one group and as early- and late-CD groups.

Antibodies to K. pneumoniae
Levels of antibody to K. pneumoniae (K50) of IgM class (Fig. 1AGo) were significantly higher in CD (mean±S.E. OD units) (0.39±0.05; t=6.6, P<0.001) and AS patients (0.24±0.03; t=6.2, P<0.001), IgG levels (Fig. 1BGo) were significantly higher in CD patients (0.32±0.05; t=5.3, P<0.001) and the AS group (0.30±0.05; t=4.7, P<0.001), and IgA levels were significantly higher (Fig. 1CGo) in CD (0.07±0.02; t=21.0, P<0.001) and AS patients (0.05±0.01; t=13.9, P<0.001) than in the healthy controls. Levels of IgM, IgG and IgA antibodies to K. pneumoniae in early and late CD were analysed in comparison with controls, and the OD units (mean±S.E.) are shown in Table 2Go. There was no difference between early or late CD patients in the mean levels of IgM, IgG and IgA class antibodies to K. pneumoniae. However, IgM anti-K. pneumoniae antibody levels were higher in early CD (t=7.5, P<0.001) and late CD (t=9.8, P<0.001) than in control subjects; levels of IgG anti-K. pneumoniae antibody were higher in early CD (t=7.7, P<0.001) and late CD (t=4.2, P<0.001) than in controls, and levels of IgA anti-K. pneumoniae antibody were higher in early CD patients (t=4.8, P<0.001) and late CD patients (t=2.9, P<0.01) than in control subjects.



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  		FIG. 1. Levels of (A) IgM, (B) IgG and (C) IgA antibodies in CD and AS patients and healthy control subjects tested against Klebsiella pneumoniae (K50). Bars show the means. The broken line represents the 95% confidence limit of the controls. Significant P values compared with controls are indicated. {square} Controls, • Crohn's disease, {blacksquare} ankylosing spondylitis.

 

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  		TABLE 2. IgM, IgG and IgA ELISA antibodies (mean±S.E. OD units) against K. pneumoniae and collagen types I, III, IV and V in early and late CD patients and healthy controls

 

Antibodies to collagen types I, III, IV and V
IgM, IgG and IgA levels to the four collagen types were measured in CD and AS patients (Fig 2Go).



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  		FIG. 2. Levels of IgM, IgG and IgA antibodies (mean±S.E.) to collagen types I (A), III (B), IV (C) and V (D) in CD and AS patients and healthy controls.

 
Compared with the individual control groups, CD patients had significantly higher levels (mean±S.E. OD units) of IgM antibodies to collagen types I (0.26±0.02; t=10.4, P<0.001), III (0.22±0.03; t=7.1, P<0.001), IV (0.1±0.01; t=9.3, P<0.001) and V (0.49±0.04; t=8.7, P<0.001), significantly higher levels of IgG antibodies to collagen types I (0.64±0.08; t=5.6, P<0.001), III (0.57±0.07; t=5.7, P<0.001), IV (0.49±0.07; t=5.1, P<0.001) and V (0.38±0.05; t=5.1, P<0.001), and significantly higher levels of IgA antibodies to collagen types I (0.08±0.01; t=4.9, P<0.001), III (0.15±0.02; t=6.2, P<0.001), IV (0.07±0.01; t=5.4, P<0.001) and V (0.1±0.02; t=5.8, P<0.001).

Compared with the healthy controls, AS patients had significantly higher levels of IgM antibodies to collagen types I (0.21±0.02; t=8.5, P<0.001), III (0.18±0.02; t=7.0, P<0.001), IV (0.07±0.01; t=5.6, P<0.001) and V (0.5±0.04; t=8.5, P<0.001), significantly higher levels of IgG antibodies to collagen types I (0.57±0.07; t=5.5, P<0.001), III (0.58±0.06; t=6.3, P<0.001), IV (0.4±0.06; t=4.2, P<0.001) and V (0.35±0.04; t=5.2, P<0.001), and significantly higher levels of IgA antibodies to collagen types I (0.07±0.02; t=3.0, P<0.001), III (0.12±0.01; t=6.3, P<0.001) and V (0.1±0.02; t=4.3, P<0.001). There was no significant difference in antibody levels of IgA antibodies to collagen type IV in the AS patients compared with the control group.

Levels of IgM, IgG and IgA antibodies to collagen types I, III, IV and V in early and late CD were analysed in comparison with controls, and the OD units (mean±S.E.) are shown in Table 2Go.

There was no difference between early and late CD in the mean levels of antibodies of the different classes to the individual collagens. Compared with the healthy control group, the group of patients with early CD had significantly higher levels of IgM antibodies to collagen types I (t=7.4, P<0.001), III (t=7.4, P<0.001), IV (t=9.6, P<0.001) and V (t=5.3, P<0.001).

The group of patients with early CD also had significantly higher values than the healthy control group for levels of IgG antibodies to collagen types I (t=4.2, P<0.001), III (t=5.0, P<0.001), IV (t=3.8, P<0.001.001) and V (t=4.7, P<0.001) and in levels of IgA antibodies to collagen types I (t=10.4, P<0.001), III (t=9.1, P<0.001), IV (t=3.1, P<0.001) and V (t=9.0, P<0.001).

The group of patients with late CD had significantly higher levels than the healthy control group for IgM antibodies to collagen types I (t=14.2, P<0.001), III (t=6.9, P<0.001), IV (t=7.7, P<0.001) and V (t=9.9, P<0.001.001), for IgG antibodies to collagen types I (t=5.2, P<0.001), III (t=5.5, P<0.001), IV (t=4.8, P<0.001) and V (t=4.6, P<0.001.001), and for IgA antibodies to collagen types I (t=5.8, P<0.001), III (t=5.4, P<0.001), IV (t=4.7, P<0.001) and V (t=5.4, P<0.001).

Patients with immunoglobulin levels above the 95% confidence limits
Numbers of patients with levels of IgM, IgG and IgA antibodies to each of the collagen types and to K. pneumoniae that were above the 95% confidence limit of the control population were determined in the disease and control groups (one-tailed test; Table 3Go) and statistical significances were assessed using the {chi}2 test with Yates’ correction (Table 4Go).


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  		TABLE 3. Numbers of early and late CD and AS patients with reactivity to collagen types I, III, IV and V and to K. pneumoniae (K50) above the 95% confidence limit (one-tailed test) of controls

 

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  		TABLE 4. {chi}2 values for numbers early and late CD and AS patients with reactivity to collagen types I, III, IV, V and K. pneumoniae (K50) above the 95% confidence limit (one-tailed test) of controls

 
Compared with the controls, significantly more early and late CD patients showed levels of antibodies to collagens I, III, IV and V and to K. pneumoniae that were above the 95% confidence. However, levels of IgG and IgA antibodies above this limit were not found in patients with early CD (Table 4Go). Furthermore, the levels of significance of the increases in the numbers of late CD and AS patients with antibody levels above the limit compared with the control group were similar for all four types of collagen, and particularly for K. pneumoniae, for all three classes of immunoglobulin, except IgG to type IV collagen in AS patients. There was no statistically significant difference between levels of antibody to any of the collagen types or to K. pneumoniae in early or late CD patients when they were stratified according to high or low CRP or to high or low ESR.

Five patients diagnosed with early CD and who also had ileal strictures, one of whom had AS and was HLA-B27-positive, had elevated levels of IgM antibodies to collagen types I, III, IV and V, whilst levels of IgG and IgA antibodies to collagen types I, III and V were above the 95% confidence limit. Furthermore, all five patients had levels of IgM and IgG antibodies to K. pneumoniae that were above the 95% confidence limits, whereas only two had IgA levels above the limits; one of these patients had AS. Four out of the five patients were male, three of whom were on prednisolone and two on Pentasa (mesalazine). Five male patients with late CD who had strictures and required surgical resections did not have levels of antibodies to any of the collagens above the 95% confidence limits, but had elevated levels of antibodies to K. pneumoniae. Furthermore, four patients (three males and one female) with late CD were diagnosed as having fistulae. All four patients had both high CRP and high ESR but did not have elevated levels of antibody to any of the collagens. However, three of these patients did have an immune response to Klebsiella above the 95% confidence limits.

Correlation coefficient analysis
There was a significant correlation between IgM anti-K. pneumoniae antibodies and IgM antibodies to type I, III and IV collagens in both early and late CD patients and to type V in early CD patients only (Table 5Go). However, there was no significant correlation between IgG anti-K. pneumoniae antibodies and type I, III, IV and V collagen antibodies in either early or late CD (Table 5Go). Levels of IgA antibodies against K. pneumoniae were also found to correlate with collagen types III and IV in early CD and with type III in late CD patients (Table 5Go).


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  		TABLE 5. Correlation coefficient (r) between K. pneumoniae IgM, IgG, IgA and antibodies to collagens type I, III, IV and V in patients and healthy controls

 
Correlation between anti-K. pneumoniae antibodies and collagen antibodies was also demonstrated in the group of AS patients. IgM and IgG anti-K. pneumoniae antibodies correlated with antibodies to collagen types I and III (Table 5Go), whereas in the case of collagen IV only IgG antibodies were found to correlate with antibacterial antibodies.

Coefficient of variation
The mean coefficients of variation for levels of IgM, IgG, IgA antibodies to collagen types I, III, IV and V and K. pneumoniae were calculated. For type I collagen, a mean coefficient of 7% was obtained for IgM, 9% for IgG and 15% for IgA. For type III collagen the coefficients were 7% for IgM, 6% for IgG and 9% for IgA. For type IV collagen they were 8% for IgM, 8% for IgG and 14% for IgA. For type V collagen they were 9% for IgM, 12% for IgG and 5% for IgA. In addition, a mean coefficient variation value for K. pneumoniae assays was calculated; the values were 6% for IgM, 10% for IgG and 8% for IgA.


   Discussion
Top
 Abstract
 Introduction
 Patients and methods
 Results
 Discussion
References
 
This is the first report showing a correlation between the immune responses to a bacterium, namely K. pneumoniae, and self-proteins, namely collagen types I, III, IV and V, in patients having either AS or CD.

This study shows that levels of IgM, IgG and IgA antibodies to collagen types I, III, IV, V and K. pneumoniae were significantly elevated in early and late CD patients compared with healthy controls. Furthermore, IgM and IgG antibodies to type I, III, IV and V collagens were found to be significantly increased in HLA-B27-positive AS patients, whereas IgA levels that were increased above the 95% confidence limits for the control group were found only for collagen types I, III and V. In addition, levels of anti-K. pneumoniae antibodies of all three classes were elevated in the AS group when compared with the control group. These findings confirm previous reports of an association between elevated levels of antibodies to K. pneumoniae in CD and AS patients [12,18]. Furthermore, this study is in agreement with recent reports of elevated IgG, IgA and IgM antibodies against collagen types I, III and IV in AS patients [6,19]. Our findings show that AS patients have immune responses to these collagens, which may be responsible for maintaining the characteristic local inflammation in AS. Type I collagen is present predominantly in tendons and bones, type III in muscle tissue and types IV and V in the basement membranes of the retina and cornea [8]. AS patients often suffer from acute anterior uveitis, which could be triggered by some of these anti-collagen antibodies [20].

Molecular mimicry has been proposed as a mechanism involved in the pathogenesis of AS. Amino acid sequence homology between HLA-B27 and the nitrogenase reductase and pullulanase enzymes of K. pneumoniae has been reported [5, 6]. Furthermore, there are similarities in amino acid sequence between the N-terminal end of the extracellular enzyme pullulanase (pulA), in the form of repeats of a tripeptide (Gly-X-Pro), and collagens of types I, III and IV [6]. In the present study there was a good correlation between IgM and IgG antibodies against K. pneumoniae and collagen types I, III and IV in AS patients. This suggests recent infection with this organism. It has been suggested that immune responses to particular serotypes of K. pneumoniae are of importance in HLA-B27-positive patients [21], class-specific IgA antibodies having a pathogenic role, since their titre correlates with disease activity [22, 23]. In this study, we used the K. pneumoniae serotype K50, which has been shown previously by our group [14] and others [21] to produce an elevated antibody response in active AS patients compared with control groups. The immune response to K. pneumoniae was found to be statistically significant in the early and late CD patients investigated in this study, when compared with healthy controls. Furthermore, IgA and IgM antibodies against both K. pneumoniae and collagen types I, III, IV and V correlated in early and late CD patients. However, high or low CRP or high or low ESR was not correlated with levels of antibodies to collagen or K. pneumoniae antibody in early and late CD patients; this may have been due to the low number of patients used in this study, or to a difference in the pathogenic mechanism involved in the initiation of disease and development of persisting chronic fibrotic lesions.

The association of AS with occult bowel disease has been well established. Gut inflammation plays a crucial role in the pathogenesis of spondylarthropathies, as ileocolonoscopic studies have demonstrated the presence of gut inflammation in 60% of AS patients [9]. In the follow-up study, 25% of those who had chronic inflammatory changes on initial biopsy had developed CD [24]. The high prevalence of IBD in spondylarthropathies confirms the hypothesis that these two disease entities have common pathogenic mechanisms.

An increased immunopathophysiological response to Klebsiella may be important in the initiation and perpetuation of intestinal inflammation in IBD, and the increased expression and secretion of pro-inflammatory cytokines such as tumour necrosis factor {alpha} plays a key role in the immunopathogenesis of CD [25]. Strictures in CD patients contain large amounts of collagen, especially types III [2] and V [1]. These findings are confirmed indirectly by our studies. Patients who had ileal strictures contained higher levels of antibodies to collagen types I, III and V than healthy controls. However, those who had undergone ileal stricture resection had normal levels of collagen antibodies.

Several independent studies have demonstrated that sulphasalazine is an effective drug in the treatment of AS and is also effective in the treatment of IBD. The drug is known to have antibiotic activity against Klebsiella, and Finnish studies have demonstrated that the titre of anti-Klebsiella antibodies decreases after treatment with sulphasalazine [26]. It has been suggested that the beneficial effect of the drug could be due to its anti-inflammatory effects on the gut wall, by normalizing its permeability and by preventing the entrance of antigens through the defective gut wall [9].

This study provides supportive evidence that there is a correlation between Klebsiella antibodies and collagen types I, III and IV antibodies in both AS and CD. In addition, Klebsiella antibodies and collagen antibodies are increased in both disease groups. Furthermore, persistently elevated immune responses to Klebsiella and to the collagens appear to be present in both early and late CD patients.

Further studies using a larger number of early and late CD patients are required in order to determine the role of K. pneumoniae and its link with collagens in the aetiopathogenesis of CD and AS.


   Acknowledgments
 
The authors gratefully acknowledge the support of the Arthritis and Rheumatism Council and the Trustees of King's College and Middlesex Hospital.


   Notes
 
Correspondence to: A. Ebringer, Division of Life Sciences, Infection and Immunity Group, King's College, Stamford Street, London, UK. Back


   References
Top
 Abstract
 Introduction
 Patients and methods
 Results
 Discussion
 References
 

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Submitted 12 November 1999; Accepted 24 July 2000


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