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Badaniami objęto lipopolisacharydy wyekstrahowane z dziewięciu klinicznych szczepów B. fragilis wyizolowanych w Polsce. Biologiczną aktywność preparatów LPS oznaczono przy użyciu fotometrycznego testu BET (poprzednio LAL). Aktywność lipopolisacharydów klinicznych szczepów B. fragilis porównano z aktywnością LPS referencyjnych szczepów B. fragilis i LPS E. coli 055:B5. Wśród lipopolisacharydów pałeczek gatunku B. fragilis największą aktywność w reakcji z odczynnikiem LAL wykazał LPS szczepu wyizolowanego z ropnia trzustki.
The aim of our study was to evoke oral tolerance with hydrolisate of spinal cord of pig used for feeding the experimental animals/ rats/After induction of tolerance animals were immunised by injection of guine pig spinal cord homogenate with Freund’s adjuvant to induce of EAE, which is an animal model of sclerosis multiplex. Clinical course was have been observed, histopatological study ,ultramicroscopic study and metalloproteinases determination in serum were done. Study of lymphocytes proliferation and level of cytokines IL-4,Il-6, Inf-gamma and TGF-beta were also performed. Clinical course of EAE after hydrolisate treatment has been milder than control. TNF in brains was decreased. Metalloproteases increased in EAE, after hydrolisate treatment were diminished by 30%. Some changes in blood brain barrier/BBB/ as opened tight junction and other changes in early phase of EAE as karioskeletal damage with vesicular structures in karioplasm, compartmentalisation of the endoplasmic reticulum in numbers of phagolisosomes, desorganisation of sheets myelin, neoangiogenesis of parenchyma of the cerebral cortex has been diminished. Mechanism of this effects is probably through active suppression involving diminishing production of IL-4 and interferon gamma as well as increasing production of Il-10 and TGF alfa. All above results indicate that mixture of neuropeptides in spinal cord hydrolisate given orally decrease immunological response to myelin antigens and gave an implication for using oral tolerance to support multiple sclerosis/ MS/ treatment.
Background: Recently has been proposed to apply a method of oral tolerance to ameliorate auto-immune reactions. The aim of this study was to use the hydrolysate of pig spinal cord proteins (mixture of neuroantigens) to induce oral tolerance in the animal model of sclerosis multiplex (SM) ñ experimental allergic encephalomyelitis (EAE). Methods: The female Lewis rats were fed with pig spinal cord hydrolysate in two doses for one week before immunization, which was induced by injection of guinea pig spinal cord homogenate. The clinical course was observed and evaluated in a fi ve grade scale. At the peak of clinical symptoms (the 13th day post immunization) the rats were sacrifi ced and the spleen removed. Splenocytes were suspended in a culture medium and placed in microculture plates. The cells were stimulated with homogenate alone, hydrolysate alone, mixture of homogenate + hydrolysate, and medium alone. The cells were cultured for seven days. Subsequently, proliferation of splenocytes was estimated by means of [3H]thymidine incorporation and expressed in cpm (average of triplicate samples). In supernatants of cultures of splenocytes the level of cytokines interferon gamma (IFN-γ), interleukin (IL)-10, IL-4, and tumor growth factor (TGF)-α was measured. Results: It was demonstrated that homogenate-induced splenocytes of hydrolysate-fed rats gave rise to low proliferation as compared to the controls used. The IFN-γ was inhibited in hydrolysate-fed animals as well as in hydrolysate-stimulated samples. Conclusion: The results show that the hydrolysate of pig spinal cord proteins has a modulatory effect on the immune reaction, particularly on the orally-induced antigen-specifi c modulation of autoimmune response. It might have a clinical implication in SM treatment.
The aim of this study was to use the hydrolysate of pig spinal cord proteins to induce oral tolerance in the animal model of sclerosis multiplex experimental allergic encephalomyelitis. The female Lewis rats were fed with hydrolysate of pig spinal cord proteins in two doses for one week before immunization, which was induced by injection of guinea pig spinal cord homogenate. At the peak of clinical symptoms (the 13th day post immunization) the rats were sacrificed and the spleen removed. Splenocytes were suspended in a culture medium and placed in microculture plates. The cells were stimulated with homogenate. The cells were cultured for seven days. Proliferation of splenocytes was estimated by means of methyl- 3H thymidine incorporation. In supernatants of cultures of splenocytes the level of cytokines INF-γ, IL-10, IL-4, and TGF-γ was measured. It was demonstrated that homogenate-induced splenocytes of hydrolysate-fed rats gave rise to low proliferation as compared to the controls used. The IFN-γ was inhibited in hydrolysate-fed animals. The hydrolysate of pig spinal cord proteins has a modulatory effect on the immune reaction, particularly on the orally-induced antigen-specific modulation of autoimmune response.
The balance between immunogenic and tolerogenic activities in human immune system strongly depends on microflora-induced pro-and anti-inflamatory activities. Lactic acid bacteria (LAB) are important components of microflora. The interactions of the different strains of LAB and the cells of immune system are largely unknown. To assess if LAB strains composition would have an effect on the cellular responses profile (proliferation, cytokines synthesis) peripheral blood mononuclear cells (PBMC) model system was used. PBMC were induced by three different strains of LAB: Lactobacillus acidophilus, Lactobacillus delbrueckii spp. bulgaricus, Bifidobacterium bifidum. Tested strains were mixed together, in combinations with each other (pairs) or alone. Both, the LAB mixture as well as the pairs and the single LAB strains induced low lymphocyte proliferation (about 10% of ConA-induced response). However, the single LAB strains and their combinations were quite different cytokines inducers. First, L. acidophilus was much stronger IFN- inducer than the LAB mixture, being a few times higher IL-12 stimulator than L. bulgaricus and B. bifidum. Second, L. bulgaricus and B. bifidum suppressed L.acidophilus-induced IFN- synthesis to the level equal to that induced by the LAB mixture, limiting IL-12 production by about 30% and 70%, respectively. Third, the LAB strains were good IL-10 and TNF-alphalfa inducers, irrespectively of their combinations used. We conclude that LAB strains’ pro or anti-inflammatory potentials are at least in part dependent on their composition. Low LAB mixture-induced IL-12 and IFN- production and relatively high IL-10 and TNF-alphalfa expression may represent cellular activities normally induced in vivo by a combined action of bacterial antigens. Their presence is important to limit pro-inflammatory reactions (via IL-10) and to provide protection against infections (via TNF-alphalfa).
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