A balanced microbiota of the gastrointestinal tract (GIT) is a prerequisite for a healthy host. The GIT microbiota in preterm infants is determined by the method of delivery and nutrition. Probiotics can improve the GIT microbiota balance and suitable animal models are required to verify their harmlessness. Preterm gnotobiotic piglets were colonized with Lactobacillus rhamnosus GG (LGG) to evaluate its safety and possible protective action against infection with an enteric pathogen, Salmonella Typhimurium (ST). Clinical signs (anorexia, somnolence, fever and diarrhea), bacterial interference and translocation, intestinal histopathology, transcriptions of claudin-1, occludin and interferon (IFN)-γ, intestinal and systemic protein levels of interleukin (IL)-8, IL-12/23 p40 and IFN-γ were compared among (i) germ-free, (ii) LGG-colonized, (iii) ST-infected and (iv) LGG-colonized and subsequently ST-infected piglets for 24 h. Both LGG and ST-colonized the GIT; LGG translocated in some cases into mesenteric lymph nodes and the spleen but did not cause bacteremia and clinical changes. ST caused clinical signs of gastroenteritis, translocated into mesenteric lymph nodes, the spleen, liver and blood, increased claudin-1 and IFN-γ transcriptions, but decreased occludin transcription and increased local and systemic levels of IL-8 and IL-12/23 p40. Previous colonization with LGG reduced ST colonization in the jejunum and translocation into the liver, spleen and blood. It partially ameliorated histopathological changes in the intestine, reduced IL-8 levels in the jejunum and plasma and IL-12/23 p40 in the jejunum. The preterm gnotobiotic piglet model of the vulnerable preterm immunocompromised infant is useful to verify the safety of probiotics and evaluate their protective effect.

• Klíčová slova
• Lactobacillus rhamnosusGG, Salmonella Typhimurium, bacterial interference, gnotobiotic piglets, preterm,
• MeSH
• bakteriální translokace MeSH
• cytokiny analýza   MeSH
• gnotobiologické modely MeSH
• Lacticaseibacillus rhamnosus * MeSH
• prasata MeSH
• předčasný porod mikrobiologie   MeSH
• probiotika farmakologie   MeSH
• proteiny těsného spoje genetika   MeSH
• Salmonella typhimurium růst a vývoj   MeSH
• střeva mikrobiologie   patologie   MeSH
• střevní mikroflóra MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH
• proteiny těsného spoje MeSH

OBJECTIVES: Alarmin high mobility group box 1 (HMGB1) is essential for correct DNA folding and transcription. It can be released from damaged cells or secreted by stimulated cells. HMGB1 has been detected in serum or plasma as a late marker of sepsis, but its suitability as a marker of sepsis has been disputed. METHODS: One-week-old germ-free piglets were orally infected/colonized with enteric bacterial pathogens (Salmonella Typhimurium or Escherichia coli O55) or with probiotic bacteria (E. coli Nissle 1917) for 24 h. The transcriptions of HMGB1, interleukin (IL)-8, tumor necrosis factor (TNF)-α, and IL-10 (quantitative reverse transcription and polymerase chain reaction), their protein levels (ELISA), and clinical state of the piglets (somnolence, anorexia, diarrhea, tachycardia, tachypnea, and tremor) were estimated. RESULTS: The piglets infected with enteric pathogens suffered from infections. HMGB1 was transcribed in the terminal ileum constitutively, regardless of any bacterial presence. In contrast, the transcription of cytokines was upregulated by virulent bacteria. HMGB1, IL-8, and TNF-α levels in the ileum were increased by both enteric pathogens, while IL-10 levels increased in E. coli O55-infected piglets only. HMGB1 significantly increased in the plasma of piglets infected with virulent E. coli only, but cytokine levels were in most cases increased by both virulent bacteria. HMGB1 and cytokine levels in ileum lavages and plasma of piglets colonized with probiotic E. coli remained comparable to those of the non-stimulated germ-free piglets. CONCLUSION: The local and systemic expression of HMGB1, its relationship to the inflammatory cytokines, and clinical findings showed HMGB1 as a suitable marker of severity of sepsis in the gnotobiotic piglet infection model.

• MeSH
• bakteriální infekce krev   imunologie   mikrobiologie   MeSH
• biologické markery krev   MeSH
• ELISA MeSH
• Escherichia coli růst a vývoj   MeSH
• gnotobiologické modely * MeSH
• ileum metabolismus   mikrobiologie   MeSH
• interleukin-10 krev   MeSH
• interleukin-8 krev   MeSH
• modely nemocí na zvířatech MeSH
• novorozená zvířata krev   imunologie   mikrobiologie   MeSH
• polymerázová řetězová reakce s reverzní transkripcí MeSH
• prasata MeSH
• protein HMGB1 * krev   MeSH
• průjem MeSH
• Salmonella typhimurium růst a vývoj   MeSH
• sepse krev   imunologie   mikrobiologie   MeSH
• stupeň závažnosti nemoci MeSH
• tachykardie MeSH
• TNF-alfa krev   MeSH
• tremor MeSH
• zánět krev   imunologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• biologické markery MeSH
• interleukin-10 MeSH
• interleukin-8 MeSH
• protein HMGB1 * MeSH
• TNF-alfa MeSH

One-week-old breast-fed miniature piglets were orally infected either with virulent LT2 strain or with a non-virulent SF1591 rough mutant of Salmonella Typhimurium for 1 d. Both microorganisms were cultivated from mesenteric lymph nodes but not from the blood of infected piglets. Interleukins (IL) 1 beta, 8, 18, tumor necrosis factor alpha (TNF-alpha) and interferon gamma (IFN-gamma) were quantified by ELISA in plasma and washes of a terminal part of the small bowel. In plasma, cytokines were mostly missing in non-infected piglets and either missing or low in infected piglets. In the gut of non-infected piglets, IL-1 beta, IL-8 and IL-18 were detected whereas TNF-alpha and IFN-gamma were mostly missing. IFN-gamma levels highly increased (p < 0.05) after infection with nonvirulent salmonellae. The variability of cytokine levels in the gut of suckling piglets is discussed.

• MeSH
• cytokiny krev   metabolismus   MeSH
• elektronová mikroskopie veterinární   MeSH
• ELISA veterinární   MeSH
• gastrointestinální nemoci imunologie   mikrobiologie   patologie   veterinární   MeSH
• ileum imunologie   mikrobiologie   patologie   ultrastruktura   MeSH
• lymfatické uzliny imunologie   mikrobiologie   patologie   ultrastruktura   MeSH
• miniaturní prasata MeSH
• nemoci prasat imunologie   mikrobiologie   patologie   MeSH
• novorozená zvířata MeSH
• prasata MeSH
• Salmonella typhimurium růst a vývoj   ultrastruktura   MeSH
• salmonelová infekce u zvířat imunologie   mikrobiologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytokiny MeSH