Gnotobiotic (GN) animals with defined microbiota allow us to study host-microbiota and microbiota-microbiota interferences. Preterm germ-free (GF) piglets were mono-associated with probiotic Bifidobacterium animalis subsp. lactis BB-12 (BB12) to ameliorate/prevent the consequences of infection with the Salmonella Typhimurium strain LT2 (LT2). Goblet cell density; expression of Toll-like receptors (TLRs) 2, 4, and 9; high mobility group box 1 (HMGB1); interleukin (IL)-6; and IL-12/23p40 were analyzed to evaluate the possible modulatory effect of BB12. BB12 prevented an LT2-induced decrease of goblet cell density in the colon. TLRs signaling modified by LT2 was not influenced by the previous association with BB12. The expression of HMGB1, IL-6, and IL12/23p40 in the jejunum, ileum, and colon and their levels in plasma were all decreased by BB12, but these changes were not statistically significant. In the colon, differences in HMGB1 distribution between the GF and LT2 piglet groups were observed. In conclusion, the mono-association of GF piglets with BB12 prior to LT2 infection partially ameliorated the inflammatory response to LT2 infection.

• Klíčová slova
• Bifidobacterium animalis subsp. lactis BB-12, Salmonella Typhimurium, Toll-like receptors, high mobility group box 1, immunodeficient host, inflammatory cytokines, intestinal barrier, mucin, tight junction proteins,
• MeSH
• Bifidobacterium animalis * MeSH
• gnotobiologické modely MeSH
• lidé MeSH
• novorozenec nedonošený MeSH
• novorozenec MeSH
• prasata MeSH
• probiotika * farmakologie   MeSH
• protein HMGB1 * MeSH
• Salmonella typhimurium MeSH
• toll-like receptory metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• protein HMGB1 * MeSH
• toll-like receptory MeSH

Cellular and humoral aspects of the immune response develop sequentially in the fetus. During the ontogeny, the pluripotent stem cells emerge and differentiate into all hematopoietic lineages. Basic questions including the identification of the first lympho-hematopoietic sites, the origin of T and B lymphocytes, the development of different subpopulations of alphabeta T, gammadelta T and B lymphocytes as well as development of innate immunity and the acquisition of full immunological capacities are discussed here for swine and compared with other species. The description of related topics such as fertilization, morphogenesis, maternal-fetal-neonatal physiology and early neonatal development are also discussed.

• MeSH
• B-lymfocyty cytologie   imunologie   metabolismus   MeSH
• buněčná imunita MeSH
• embryo savčí imunologie   MeSH
• hematopoetické kmenové buňky cytologie   imunologie   MeSH
• hematopoéza imunologie   MeSH
• imunitní systém embryologie   imunologie   MeSH
• lymfopoéza MeSH
• maternofetální výměna látek imunologie   MeSH
• morfogeneze imunologie   MeSH
• placentace imunologie   MeSH
• prasata embryologie   imunologie   virologie   MeSH
• přirozená imunita MeSH
• T-lymfocyty cytologie   imunologie   metabolismus   MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

In this study, two stable, rough, streptomycin-sensitive Salmonella mutants with different types of genetic defects were used to colonize groups of germ-free (GF) piglets. The lipopolysaccharide (LPS) of Salmonella typhimurium SF 1591 was of the Ra chemotype (complete core), whereas the LPS of the S. minnesota mR 595 deep-rough mutant contained only lipid A and 2-keto-3-deoxyoctulosonic acid (Re chemotype). Both strains readily colonized the intestinal tracts of GF piglets and were stable during the whole experiment. All animals survived, and only transient fever was observed in some piglets colonized with the SF 1591 strain. Finally, streptomycin and virulent, smooth, streptomycin-resistant S. typhimurium LT2 were administered perorally 1 week later. All piglets colonized previously with the deep-rough mutant mR 595 died of sepsis, in contrast to piglets infected with the LT2 strain and colonized with the SF 1591 mutant, all of which survived. This difference is explained by the penetration of the mesenteric lymph nodes, spleen, and liver by great numbers of live bacteria in the latter case, resulting in prominent systemic and local immune responses. On the other hand, live bacteria were found only rarely in the mesenteric lymph nodes of animals colonized with the mR 595 strain and a negligible antibody response was observed.

• MeSH
• gnotobiologické modely * MeSH
• miniaturní prasata MeSH
• mutace * MeSH
• prasata MeSH
• Salmonella genetika   patogenita   MeSH
• salmonelová infekce u zvířat mikrobiologie   MeSH
• virulence genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Pig fetuses, colostrum-deprived newborns and germ-free (GF) piglets, animals in which B-cell development is not influenced by maternal regulatory factors, were employed to study the occurrence and specificity of natural antibodies (NAb). Serum immunoglobulins of all isotypes were found in 44-day-old fetuses (the gestation period in pigs lasts 114 days) and their level, with predominating IgM, was increased during fetal ontogeny. In sera of fetuses at the end of embryonic life as well as of newborns and older GF piglets, antibody activity against autoantigens (thyroglobulin, hormones, ssDNA), phylogenetically conserved proteins (myosin), haptens (trinitrophenyl; TNP) and bacterial components (Escherichia coli O86, tetanic anatoxin) was detected by enzyme-linked immunosorbent assay. The antigen-biding activity of IgM NAb increased after isolation of the serum immunoglobulins on a Staphylococcus Protein A (SPA)-Sepharose column. IgM reactivity similar to that detected in serum was found in supernatants from polyclonally stimulated cultures of spleen of 8- and 12-day-old GF piglets. Pig fetal liver IgM+ B cells, which were able to produce IgM after polyclonal stimulation, did not express the CD5 molecule. Our results indicate that pig preimmune repertoire is comparable to that described in humans and mice, although in contrast to these species pig B-1 cells do not express CD5.

• MeSH
• antigeny CD5 analýza   MeSH
• B-lymfocyty imunologie   MeSH
• buněčné kultury MeSH
• gnotobiologické modely MeSH
• imunoglobulin M biosyntéza   MeSH
• imunoglobulinové izotypy krev   MeSH
• imunoglobuliny biosyntéza   MeSH
• játra embryologie   imunologie   MeSH
• novorozená zvířata MeSH
• plod imunologie   MeSH
• prasata imunologie   MeSH
• specificita protilátek MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD5 MeSH
• imunoglobulin M MeSH
• imunoglobulinové izotypy MeSH
• imunoglobuliny MeSH

The aim of this study was to investigate spontaneous immunoglobulin production and a pattern of isotype switching by thymic B lymphocytes (TBL) as compared with cells isolated from spleen during early ontogeny using a pig model in which B-cell development is not influenced by maternal regulatory factors. A sensitive ELISPOT assay was therefore employed to detect immunoglobulins in pig fetuses, colostrum-deprived germ-free (GF) piglets as well as conventionally (CONV) reared pigs. The first spontaneously immunoglobulin-secreting cells in the thymus were detected in 67-day-old fetuses (the length of gestation period in pigs is 114 days), their number increasing during fetal ontogeny. In contrast to fetal splenic cells, which secrete exclusively IgM, fetal thymic immunoglobulin-secreting cells were determined to undergo spontaneous isotype switching to IgG and IgA. In 28-day-old GF piglets and 3-month-old CONV pigs the number of thymic immunoglobulin-secreting cells of all isotypes was comparable to the number of thymic immunoglobulin-secreting cells detected in the newborn thymus. Considerable augmentation of IgG and IgA production by splenic immunoglobulin-secreting cells in CONV pigs was observed as compared to GF newborns and GF piglets, in which IgG- and IgA-secreting cells were detected occasionally. Our results indicate that TBL represent the first B-cell population in early fetal ontogeny spontaneously undergoing isotype switching to IgG and IgA; in the postnatal period the TBL population does not appear to be influenced by external antigenic stimuli of conventional microflora.

• MeSH
• B-lymfocyty imunologie   MeSH
• ELISA MeSH
• gestační stáří MeSH
• gnotobiologické modely * MeSH
• imunoglobulin A biosyntéza   MeSH
• imunoglobulin G biosyntéza   MeSH
• imunoglobulin M biosyntéza   MeSH
• imunoglobulinové izotypy biosyntéza   MeSH
• miniaturní prasata embryologie   růst a vývoj   imunologie   MeSH
• prasata MeSH
• přesmyk imunoglobulinových tříd MeSH
• slezina růst a vývoj   imunologie   MeSH
• thymus embryologie   růst a vývoj   imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• imunoglobulin A MeSH
• imunoglobulin G MeSH
• imunoglobulin M MeSH
• imunoglobulinové izotypy MeSH

Immunoglobulin (Ig) response to different polyclonal B-cell activators was measured by ELISA in cell culture media of thymocytes, splenocytes and liver cells isolated from pig fetuses, 8-d-old germ-free piglets and conventionally reared pigs. Both in fetal and in postnatal life polyclonally stimulated lymphocytes were found to produce predominantly the IgM isotype; the first IgM formation was detected in 50-d-old fetal liver (gestation in pigs lasts 114 d). Surprisingly, 73-d-old fetal thymic cells were shown to be induced to Ig synthesis and secretion. In contrast to splenocytes of the same age, which secreted exclusively IgM, fetal thymocytes produced IgM, IgG and IgA. Polyclonally stimulated splenic cells as compared with thymic cells started to produce IgA later in fetal ontogeny, whereas the IgG response was not detectable in splenic cell culture media during the whole embryonal development and appeared only after birth. The earliest and the highest Ig stimulation was found after cultivation of lymphocytes with Nocardia delipidated cell mitogen. Interestingly, the moderate stimulatory effect of 65-kDa heat shock protein (Hsp-65) in polyclonal IgM response of fetal splenocytes was observed. We showed that thymic B lymphocytes represent probably the first maturing B cell population detectable in fetal life, which is able to differentiate after polyclonal stimulation into IgM as well as IgA and IgG producing cells.

• MeSH
• aktivace lymfocytů * MeSH
• B-lymfocyty účinky léků   imunologie   MeSH
• bakteriální proteiny * MeSH
• chaperon hsp60 MeSH
• chaperoniny imunologie   MeSH
• ELISA MeSH
• gnotobiologické modely imunologie   MeSH
• játra cytologie   embryologie   růst a vývoj   imunologie   MeSH
• lipopolysacharidy imunologie   MeSH
• mitogeny líčidla amerického imunologie   MeSH
• mitogeny imunologie   MeSH
• organické látky MeSH
• orgánové kultury - kultivační techniky MeSH
• plod imunologie   MeSH
• prasata embryologie   růst a vývoj   imunologie   MeSH
• slezina cytologie   imunologie   MeSH
• thymus cytologie   embryologie   růst a vývoj   imunologie   MeSH
• tvorba protilátek * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• bakteriální proteiny * MeSH
• chaperon hsp60 MeSH
• chaperoniny MeSH
• heat-shock protein 65, Mycobacterium MeSH Prohlížeč
• lipopolysacharidy MeSH
• mitogeny líčidla amerického MeSH
• mitogeny MeSH
• Nocardia delipidated cell mitogen MeSH Prohlížeč
• organické látky MeSH