Association of germfree mice with intestinal microfloras obtained from 'normal' mice

Clinical and Experimental Immunology

et al. 1992

SUMMARYStreptococcal cell wall (SCW)-induced arthritis is a chronic, erosive polyarthritis that can be induced in susceptible Lewis rats by one i,p, injection of an aqueous, sterile suspension of SCW, F344 rats are resistant to chronic joint inflammation. Our previous studies showed a correlation between susceptibility to SCW-induced arthritis and the ability to mount SCW-specific T cell responses, suggesting tolerance to SCW as a putative mechanism. Here we prevented the induction of tolerance to bacterial epitopes in F344 rats by using them germ-free and analysed susceptibility to arthritis subsequently. In addition, we conventionalized germ-free F344 rats at different times before induction of arthritis. Our results show that germ-free F344 rats are susceptible to SCW-induced arthritis with a similar severity, chronicity, incidence and onset as Lewis rats. Moreover, T cells isolated from germ-free F344 rats were able to respond to SCW, Conventionalization dramatically moderates arthritis and makes T cells unresponsive to SCW again. Thus, in normal rats (F344) a state of tolerance to arthritogenic epitopes is induced (neonatally) and maintained through life by the bacterial flora, resulting in resistance to bacterium-induced artritides. In arthritis-prone (Lewis) rats, this tolerance is deficient and/or easily broken.

Section: Ratsmentioning

confidence: 99%

Gut flora induces and maintains resistance against streptococcal cell wall-induced arthritis in F344 rats

Broek

Bruggen

Clinical and Experimental Immunology

et al. 1992

“…The villus: crypt ratio (VCR) of the ileum was determined as described earlier (Koopman et al, 1981). IgA-producing cells in the jejunum and ileum wall were measured as described earlier (Koopman et al, 1982a).…”

Section: Relative Caecal Weight (Rcw)mentioning

confidence: 99%

“…These were measured as described earlier (Koopman et al, 1981). The results are based on a pooled sample obtained from 8 mice.…”

Section: Volatile Fatty Acids (Vfa) and Non-volatile Fatty Acids (Na)mentioning

confidence: 99%

“…In earlier experiments we tried to achieve 'normalization' of GF mice by inoculating combinations of bacterial strains, isolated from the intestines of 'normal' mice (Welling, Groen, Tuinte, Koopman & Kennis, 1980;Koopman, Welling, Huybregts, Mullink & Prins, 1981;Koopman et al, 1983a). These trials were unsuccessful, only partial 'normalization' being achieved.…”

mentioning

confidence: 99%

See 1 more Smart Citation

'Normalization' of germfree mice with anaerobically cultured caecal flora of 'normal' mice

Kennis²,

Mullink³

et al. 1984

Lab Anim

Self Cite

Germfree (GF) mice were inoculated with a cultured flora from 10-1, 10-3, 10-5, and 10-7 dilutions of caecal contents from a 'normal' mouse. GF mice associated with a flora of a 'normal' mouse served as controls. The following intestinal parameters were determined: Colonization resistance (CR), Relative caecal weight (RCW), villus:crypt ratio (jejunum and ileum), IgA-producing cells (jejunum and ileum), ß-aspartyl glycine (faeces), volatile and non-volatile fatty acids (caecum) and bile acids (faeces). Only the 10-1 culture was able to induce similar changes in the GF mice to a 'normal' flora. The GF + 10-5 and GF + 10-7 groups deviated markedly from the controls while the GF + 10-3 group showed in general intermediate values between GF + SPF and GF + 10-1 on the one hand and GF + 10-5 and GF + 10-7 on the other hand. ß-aspartyl glycine was present only in the GF + 10-7 group. Scanning electron microscopy (SEM) of ileal contents revealed segmented filamentous organisms in the ileum of controls and the GF + 10-1 group. The faecal flora consisted mainly of fusiform organisms. In the faeces of the 10-5 and 10-7 groups increasing amounts of non-bacterial matter were found, while in the faeces of the other groups virtually only bacteria were seen.

“…The caecum and parts of the ileum (± 5 cm) of both animals were homogenized in broth as described earlier (Koopman et al, 1983). From this homogenate a 10-2 dilution was made and 0'1 ml of this was spread on the surface of agar plates (Koopman et al, 1983).…”

Section: Preparation Of the Donor Microfloramentioning

confidence: 99%

The use of a rat-derived microflora for providing colonization resistance in SPF rats

Heidt

Kennis³

et al. 1990

Lab Anim

Self Cite

SummaryTo obtain a suitable species-specific microflora for a new rat SPF -unit, germ-free WAG/Rij rats were associated with a flora derived originally from selectively decontaminated Cpb : WU (Wistar) rats. Caecal and ileal contents of these rats had been cultured anaerobically (37°C) for 7 days and harvested. This cultured flora was given to germ-free Cpb : SE (Swiss) mice, which were kept in an isolator system and acted as a source of the flora to associate germ-free Wag/Rij rats.In these associated rats, several parameters indicative of the 'quality' of the intestinal micro flora were investigated and compared to those in rats with a mouse derived anaerobic microflora. Parameters included relative caecal weight, colonization resistance and the concentration of faecal bile acids. The cultured rat-derived microflora normalized the observed intestinal parameters better than the mouse derived microflora, and provided better colonization resistance.We conclude that culturing of intestinal contents of selectively decontaminated animals can be a useful way to obtain a species-specific donor-micro flora which can be used to start new SPF units.