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.

• Keywords
• 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
• Germ-Free Life MeSH
• Humans MeSH
• Infant, Premature MeSH
• Infant, Newborn MeSH
• Swine MeSH
• Probiotics * pharmacology   MeSH
• HMGB1 Protein * MeSH
• Salmonella typhimurium MeSH
• Toll-Like Receptors metabolism   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Infant, Newborn MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• HMGB1 Protein * MeSH
• Toll-Like Receptors 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-Lymphocytes cytology   immunology   metabolism   MeSH
• Immunity, Cellular MeSH
• Embryo, Mammalian immunology   MeSH
• Hematopoietic Stem Cells cytology   immunology   MeSH
• Hematopoiesis immunology   MeSH
• Immune System embryology   immunology   MeSH
• Lymphopoiesis MeSH
• Maternal-Fetal Exchange immunology   MeSH
• Morphogenesis immunology   MeSH
• Placentation immunology   MeSH
• Swine embryology   immunology   virology   MeSH
• Immunity, Innate MeSH
• T-Lymphocytes cytology   immunology   metabolism   MeSH
• Pregnancy MeSH
• Animals MeSH
• Check Tag
• Pregnancy MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Review 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
• Germ-Free Life * MeSH
• Swine, Miniature MeSH
• Mutation * MeSH
• Swine MeSH
• Salmonella genetics   pathogenicity   MeSH
• Salmonella Infections, Animal microbiology   MeSH
• Virulence genetics   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't 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
• CD5 Antigens analysis   MeSH
• B-Lymphocytes immunology   MeSH
• Cell Culture Techniques MeSH
• Germ-Free Life MeSH
• Immunoglobulin M biosynthesis   MeSH
• Immunoglobulin Isotypes blood   MeSH
• Immunoglobulins biosynthesis   MeSH
• Liver embryology   immunology   MeSH
• Animals, Newborn MeSH
• Fetus immunology   MeSH
• Swine immunology   MeSH
• Antibody Specificity MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• CD5 Antigens MeSH
• Immunoglobulin M MeSH
• Immunoglobulin Isotypes MeSH
• Immunoglobulins 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-Lymphocytes immunology   MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Gestational Age MeSH
• Germ-Free Life * MeSH
• Immunoglobulin A biosynthesis   MeSH
• Immunoglobulin G biosynthesis   MeSH
• Immunoglobulin M biosynthesis   MeSH
• Immunoglobulin Isotypes biosynthesis   MeSH
• Swine, Miniature embryology   growth & development   immunology   MeSH
• Swine MeSH
• Immunoglobulin Class Switching MeSH
• Spleen growth & development   immunology   MeSH
• Thymus Gland embryology   growth & development   immunology   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH
• Names of Substances
• Immunoglobulin A MeSH
• Immunoglobulin G MeSH
• Immunoglobulin M MeSH
• Immunoglobulin Isotypes 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
• Lymphocyte Activation * MeSH
• B-Lymphocytes drug effects   immunology   MeSH
• Bacterial Proteins * MeSH
• Chaperonin 60 MeSH
• Chaperonins immunology   MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Germ-Free Life immunology   MeSH
• Liver cytology   embryology   growth & development   immunology   MeSH
• Lipopolysaccharides immunology   MeSH
• Pokeweed Mitogens immunology   MeSH
• Mitogens immunology   MeSH
• Organic Chemicals MeSH
• Organ Culture Techniques MeSH
• Fetus immunology   MeSH
• Swine embryology   growth & development   immunology   MeSH
• Spleen cytology   immunology   MeSH
• Thymus Gland cytology   embryology   growth & development   immunology   MeSH
• Antibody Formation * MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Bacterial Proteins * MeSH
• Chaperonin 60 MeSH
• Chaperonins MeSH
• heat-shock protein 65, Mycobacterium MeSH Browser
• Lipopolysaccharides MeSH
• Pokeweed Mitogens MeSH
• Mitogens MeSH
• Nocardia delipidated cell mitogen MeSH Browser
• Organic Chemicals MeSH