Investigation of association of mycobacteria with inflammatory bowel disease by nucleic acid hybridization

Abstract

We used DNA-DNA hybridization to characterize a mycobacterial isolate, strain Linda, that was obtained from a patient with Crohn's disease and that has been reported to cause ileitis in experimental animals. We also investigated the association of this mycobacterium with Crohn's disease. Our results identified Mycobacterium strain Linda as a strain of Mycobacterium paratuberculosis, the etiologic agent of Johne's disease, a disease of ruminants that has some superficial resemblance to Crohn's disease. Sequences that hybridized with strain Linda DNA probes were detected in DNA extracted from human intestinal tissues from patients with Crohn's disease, ulcerative colitis, and noninflammatory bowel disease. These hybridizing DNA sequences were more prevalent in the muscle layers than in the intestinal mucosa, making it unlikely that they represented DNA from bacterial contaminants in the intestinal lumen. Measurement of the melting temperatures of the DNA-DNA hybrids formed between strain Linda probes and tissue DNAs indicated that the related sequences detected were of mycobacterial origin but were not identical to each other or to strain Linda DNA. These results do not support the proposed specific relationship between Mycobacterium strain Linda and Crohn's disease. The possible etiologic role of mycobacteria in Crohn's disease is discussed.

Intestinal multiplication of Mycobacterium paratuberculosis in athymic nude gnotobiotic mice

“…This study was performed with a strain of M. paratuberculosis isolated from a Crohn's disease patient (strain Linda, ATCC 43015) to evaluate the in vivo virulence of M. paratuberculosis of human origin. Although M. paratuberculosis (9,11,38), other unidentified mycobacteria (4, 10), and poorly described transmissible agents (5,17), have been isolated from a number of Crohn's disease patients, their role in the etiology of human inflammatory bowel disease remains controversial (18,20,36,40,41). Crohn's disease and other inflammatory bowel diseases are heterogeneous syndromes that may vary greatly in their clinical presentation from nearly complete remission to severe debilitating disease (24).…”

Mycobacterium aviumsubspeciesparatuberculosis –an important food borne pathogen of high public health significance with special reference to India: an update

“…He therefore speculated that MAP could be a prospective etiological agent for the complications observed in human patients (Dalziel 1989). Chiodini et al (1984) revived the association of MAP with CD and reported the isolation of un-characterized mycobacteria from tissues of three CD patients and Yoshimura et al (1987) anticipated that the pathogen present in a cell-wall deficient form was characterized as MAP. In another study, MAP was directly visualized by light microscopy in small numbers in the intestines of CD patients by ZN staining (Jeyanathan et al 2007), while other pathogens (E. coli, Enterococcus spp, Clostridium perfringens, Proteus spp and Bacteroides fragilis) have also been detected in the intestines of patients with CD (De Hertogh et al 2008).…”

Evaluation of Surgical Tissue From Patients with Crohnʼs Disease for the Presence of Mycobacterium avium Subspecies paratuberculosis DNA by In Situ Hybridization and Nested Polymerase Chain Reaction

“…8 This was followed by several studies, which resulted in mixed results in which MAP or MAP DNA presence ranged from 0 to 100%. [9][10][11][12][13][14][15][16][17] In 2001, the first susceptibility gene for CD, Nod2 (nucleotidebinding oligomerization domain), was identified. 18,19 Mutations in Nod2 gene have been shown to be associated with CD, suggesting a role for intracellular pathogen-host interactions in the etiology of CD.…”