Developmental pathways for B cell lymphogenesis are sufficiently known only in mice and humans. However, both of these species rearrange immunoglobulin heavy chains (IgH) before light chains (IgL) while IgL precedes IgH rearrangement in swine. We demonstrate here that this reversed order of rearrangements have some concealed consequences: (1) we confirmed that although IgLκ rearrangement is initial, most IgLλ+ B cells are generated earlier and before IgH rearrangements, while most IgLκ+ B cells later and after IgH rearrangements, (2) the second IgLκ rearrangement can occur after IgLλ rearrangement, (3) early formed B cells bear only single in-frame IgH rearrangements, (4) many IgLκ+ B cells carry IgLλ rearrangements that can be productive and occurring on both alleles in one cell, and (5) although VpreB and λ5 genes are present in swine, they are preferentially expressed in non-B cells. In summary, our findings reveal that swine use an alternative B cell developmental pathway as compared to mice and humans.

• MeSH
• B-lymfocyty fyziologie   MeSH
• buněčná diferenciace MeSH
• genová přestavba B-lymfocytů MeSH
• imunoglobuliny - kappa-řetězce genetika   MeSH
• imunoglobuliny - lambda-řetězce genetika   MeSH
• kultivované buňky MeSH
• lidé MeSH
• myši MeSH
• prasata imunologie   MeSH
• receptory antigenů B-buněk genetika   MeSH
• těžké řetězce imunoglobulinů genetika   MeSH
• transkriptom MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

High mobility group box 1 (HMGB1) is a DNA-binding nuclear protein that can be actively secreted by immune cells after different immune stimuli or passively released from cells undergoing necrosis. HMGB1 amplifies inflammation, and its hypersecretion contributes to multiple organ dysfunction syndrome and death. We tested possible immunomodulatory effect of commensal Lactobacillus amylovorus (LA), Lactobacillus mucosae (LM) or probiotic Escherichia coli Nissle 1917 (EcN) in infection of gnotobiotic piglets with Salmonella Typhimurium (ST). Transcription of HMGB1 and Toll-like receptors (TLR) 2, 4, and 9 and receptor for advanced glycation end products (RAGE), TLR4-related molecules (MD-2, CD14, and LBP), and adaptor proteins (MyD88 and TRIF) in the ileum and colon were measured by RT-qPCR. Expression of TLR4 and its related molecules were highly upregulated in the ST-infected intestine, which was suppressed by EcN, but not LA nor LM. In contrast, HMGB1 expression was unaffected by ST infection or commensal/probiotic administration. HMGB1 protein levels in the intestine measured by ELISA were increased in ST-infected piglets, but they were decreased by previous colonization with E. coli Nissle 1917 only. We conclude that the stability of HMGB1 mRNA expression in all piglet groups could show its importance for DNA transcription and physiological cell functions. The presence of HMGB1 protein in the intestinal lumen probably indicates cellular damage.

• MeSH
• Escherichia coli imunologie   MeSH
• gnotobiologické modely imunologie   MeSH
• Lactobacillus acidophilus imunologie   MeSH
• prasata * imunologie   mikrobiologie   MeSH
• probiotika * MeSH
• protein HMGB1 imunologie   MeSH
• Salmonella typhimurium imunologie   MeSH
• signální transdukce imunologie   MeSH
• střeva imunologie   mikrobiologie   MeSH
• toll-like receptor 4 imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Using a porcine model, we describe Melanoma-Associated CD4+CD8hi T-lymphocytes (MATL) in peripheral blood that increase during melanoma regression. These MATL possess the CD4+CD8hi phenotype and they have their direct counterparts in Tumor Infiltrating Lymphocytes (TIL) isolated from melanoma loci. Both MATL and CD4+CD8hi TIL have a similar expression of selected markers indicating that they represent effector/memory αβ T-cell subset. Moreover, although TIL also contain CD4-CD8+ T-cells, only CD4+CD8hi TIL expand during melanoma regression. Importantly, TIL isolated from different pigs and different melanoma loci among the same pig have similar composition of CD4/CD8 subsets, indicating that the composition of the MATL and TIL compartment is identical. Analysis of sorted cells from regressing pigs revealed a unique MATL subpopulation with mono-specific T-cell receptor that was further analyzed by sequencing. These results indicate that pigs regressing melanomas possess a characteristic population of recirculating T-cells playing a role in tumor control and regression.

• MeSH
• antigeny CD4 metabolismus   MeSH
• antigeny CD8 metabolismus   MeSH
• cytotoxicita imunologická MeSH
• experimentální nádory imunologie   MeSH
• imunofenotypizace MeSH
• kultivované buňky MeSH
• lidé MeSH
• melanom imunologie   MeSH
• modely nemocí na zvířatech MeSH
• prasata imunologie   MeSH
• receptory antigenů T-buněk alfa-beta metabolismus   MeSH
• spontánní regrese nádoru MeSH
• T-lymfocyty imunologie   MeSH
• tumor infiltrující lymfocyty imunologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Porcine thymus contains three independent populations of cells that have rearranged immunoglobulin heavy chain VDJH genes. The first population can be found exclusively in medulla and it consists of existing mature B cells and plasma cells. The second consists of developing B cells characterized by the presence of selected VDJH rearrangement, similar to B cell lymphogenesis in the bone marrow. The third population is entirely unaffected by selection mechanism for productive VDJH rearrangement and represents T lineage cells that rearrange immunoglobulin genes. Transcription of unselected VDJH repertoire is not allowed in T cells. Sequence analysis of unselected VDJH repertoire from T cells also revealed important consequences for B cell lymphogenesis and selection of B cell repertoire. As far as we know, this is the first evidence that some species completely rearrange VDJH genes in T cells. Our results also support the finding that B cells actively develop in the thymus.

• MeSH
• B-lymfocyty imunologie   MeSH
• druhová specificita MeSH
• geny pro těžké řetězce imunoglobulinů genetika   MeSH
• lidé MeSH
• plod imunologie   MeSH
• podskupiny lymfocytů imunologie   MeSH
• prasata genetika   růst a vývoj   imunologie   MeSH
• T-lymfocyty imunologie   MeSH
• thymus růst a vývoj   imunologie   MeSH
• V(D)J rekombinace genetika   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

We describe the domestication of the species, explore its value to agriculture and bioscience, and compare its immunoglobulin (Ig) genes to those of other vertebrates. For encyclopedic information, we cite earlier reviews and chapters. We provide current gene maps for the heavy and light chain loci and describe their polygeny and polymorphy. B-cell and antibody repertoire development is a major focus, and we present findings that challenge several mouse-centric paradigms. We focus special attention on the role of ileal Peyer's patches, the largest secondary lymphoid tissues in newborn piglets and a feature of all artiodactyls. We believe swine fetal development and early class switch evolved to provide natural secretory IgA antibodies able to prevent translocation of bacteria from the gut while the bacterial PAMPs drive development of adaptive immunity. We discuss the value of using the isolator piglet model to address these issues.

• MeSH
• B-lymfocyty MeSH
• geny pro imunoglobuliny * MeSH
• ileum MeSH
• myši MeSH
• novorozená zvířata MeSH
• Peyerovy pláty MeSH
• prasata genetika   imunologie   MeSH
• tvorba protilátek * MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The ileal Peyers patches (IPP) of newborn germfree (GF) piglets were isolated into blind loops and the piglets colonized with a defined probiotic microflora. After 5 weeks, IgA levels in the intestinal lavage (IL) of loop piglets remained at GF levels and IgM comprised ∼70% while in controls, IgA levels were elevated 5-fold and comprised ∼70% of total Igs. Loop piglets also had reduced serum IgA levels suggesting the source of serum IgA had been interrupted. The isotype profile for loop contents was intermediate between that in the IL of GF and probiotic controls. Surprisingly, colonization alone did not result in repertoire diversification in the IPP. Rather, colonization promoted pronounced proliferation of fully switched IgA(+)IgM(-) B cells in the IPP that supply early, non-diversified "natural" SIgA antibodies to the gut lumen and a primary IgA response in serum.

• MeSH
• aktivace lymfocytů MeSH
• B-lymfocyty fyziologie   MeSH
• buněčná diferenciace MeSH
• gnotobiologické modely MeSH
• ileum imunologie   MeSH
• imunoglobulin A sekreční genetika   MeSH
• imunoglobulin M genetika   MeSH
• imunologická paměť MeSH
• kultivované buňky MeSH
• novorozená zvířata MeSH
• Peyerovy pláty imunologie   MeSH
• prasata imunologie   MeSH
• přesmyk imunoglobulinových tříd MeSH
• probiotika aplikace a dávkování   MeSH
• proliferace buněk MeSH
• rozmanitost protilátek MeSH
• střevní mikroflóra imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

Pig is a domestic species of major importance in the agro-economy and in biomedical research. Mononuclear phagocytes (MNP) are organized in subsets with specialized roles in the orchestration of the immune response and new tools are awaited to improve MNP subset identification in the pig. We cloned pig CD11c cDNA and generated a monoclonal antibody to pig CD11c which showed a pattern of expression by blood and skin MNP subsets similar to humans. We also developed a porcine XCL1-mCherry dimer which specifically reacted with the XCR1-expressing dendritic cell subset of the type 1 lineage in blood and skin. These original reagents will allow the efficient identification of pig MNP subsets to study their role in physiological and pathological processes and also to target these cells in novel intervention and vaccine strategies for veterinary applications and preclinical evaluations.

• MeSH
• antigeny CD11c imunologie   metabolismus   MeSH
• dendritické buňky fyziologie   MeSH
• fagocyty imunologie   MeSH
• imunologické testy metody   MeSH
• klonování DNA MeSH
• kůže metabolismus   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• monoklonální protilátky metabolismus   MeSH
• mononukleární fagocytární systém * MeSH
• prasata imunologie   MeSH
• receptory spřažené s G-proteiny imunologie   metabolismus   MeSH
• veterinární lékařství MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

A course and a site of B cell development in swine are not firmly known. In this study, we show that B cell lymphogenesis is located in the bone marrow (BM). According to expression of MHC class II (MHC-II), CD2, CD21, CD25, CD45RC, CD172a, swine workshop cluster (identification number) (SWC) 7, and μHC, porcine BM cells were resolved into seven subsets representing sequential stages of development. Profile of rearrangement-specific products and transcripts from sorted BM cells confirmed the proposed developmental pathway. The same developmental pathway was further proven by analysis of selection for productive rearrangements in Ig H chains and also by cultivation studies. Cultivation also showed that earliest precursors with incomplete DJ rearrangements can still revert their B cell differentiation and develop along myeloid lineage, whereas this is impossible for later developmental stages. Proliferation and the apoptotic potential of individual developmental stages as well as critical checkpoints were also identified. Colocalization experiments showed early colocalization of MHC-II/CD2/CD172a is replaced by colocalization of MHC-II/CD2/CD21/SWC7/IgM in immature cells, whereas CD25 and CD45RC did not colocalize with any other studied molecules. In this study, we also finally prove that the BM in pigs is fully functional in adult animals and that B lymphogenesis occurs there throughout life. To our knowledge, this is the first study showing a course and a direct site of B cell lymphogenesis in swine.

• MeSH
• buněčná diferenciace MeSH
• buňky kostní dřeně cytologie   imunologie   MeSH
• CD antigeny genetika   imunologie   MeSH
• gnotobiologické modely MeSH
• histokompatibilita - antigeny třídy II genetika   imunologie   MeSH
• hysterektomie MeSH
• imunofenotypizace MeSH
• novorozená zvířata MeSH
• podskupiny B-lymfocytů cytologie   imunologie   MeSH
• prasata imunologie   MeSH
• primární buněčná kultura MeSH
• těžké řetězce imunoglobulinů genetika   imunologie   MeSH
• vývojová regulace genové exprese imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The effects of live yeast Saccharomyces cerevisiae (strain CNCM I-4407, 10(10) cfu/g; Actisaf; Lesaffre Feed Additives, Marcq-en-Baroeul, France) on the severity of diarrhea, immune response, and growth performance in weaned piglets orally challenged with enterotoxigenic Escherichia coli (ETEC) strain O149:K88 were investigated. Live yeast was fed to sows and their piglets in the late gestation, suckling, and postweaning periods. Sows were fed a basal diet without (Control; n = 2) or with (Supplemented; n = 2) 1 g/kg of live yeast from d 94 of gestation and during lactation until weaning of the piglets (d 28). Suckling piglets of the supplemented sows were orally treated with 1 g of live yeast in porridge carrier 3 times a week until weaning. Weaned piglets were fed a basal starter diet without (Control; n = 19) or with (Supplemented; n = 15) 5 g of live yeast/kg feed for 2 wk. Significantly lower daily diarrhea scores (P < 0.05), duration of diarrhea (P < 0.01), and shedding of pathogenic ETEC bacteria (P < 0.05) in feces was detected in the supplemented piglets. Administration of live yeast significantly increased (P < 0.05) IgA levels in the serum of piglets. Evidence indicates that decreased infection-related stress and severity of diarrhea in yeast-fed weaned piglets positively affected their growth capacity in the postweaning period (P < 0.05). The results suggest that dietary supplementation with live yeast S. cerevisiae to sows and piglets in the late gestation, suckling, and postweaning periods can be useful in the reduction of the duration and severity of postweaning diarrhea caused by ETEC.

• MeSH
• dieta veterinární   MeSH
• enterotoxigenní Escherichia coli klasifikace   MeSH
• fyziologie výživy v mateřství MeSH
• fyziologie výživy zvířat MeSH
• imunoglobulin A krev   MeSH
• infekce vyvolané Escherichia coli prevence a kontrola   veterinární   MeSH
• krmivo pro zvířata MeSH
• nemoci prasat etiologie   prevence a kontrola   MeSH
• prasata růst a vývoj   imunologie   MeSH
• průjem etiologie   prevence a kontrola   veterinární   MeSH
• Saccharomyces cerevisiae * MeSH
• těhotenství MeSH
• zvířata MeSH
• Check Tag
• těhotenství MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky kontrolované MeSH
• práce podpořená grantem MeSH

Monoclonal antibodies IAH-CC51, BB6-11C9.6 and B-Ly4 are routinely used to detect CD21 orthologue on the surface of porcine B lymphocytes. Cross-reactive studies show that IAH-CC51 and B-Ly4 recognize only a portion of B cells that are positive for pan-specific BB6-11C9.6. This indicates that CD21 is always present on all mature B cells but can be expressed in at least two differential forms, and these were assigned as CD21(a) and CD21(b). We used IAH-CC51 together with anti-CD2 to define four subpopulations of B cells. Ontogenetic and in vitro culture studies, analysis of cell size, expression of CD11b and class-switched phenotype together with measurement of proliferation and cell death, revealed that these subsets represent distinct populations. Phenotypic and functional features collectively suggest that CD21(b+) B cells are less mature than CD21(b-). The present work is the first to show that distinct subsets of mature B cells can express differential forms of CD21.

• MeSH
• apoptóza MeSH
• epitopy imunologie   MeSH
• kultivované buňky MeSH
• monoklonální protilátky imunologie   MeSH
• podskupiny B-lymfocytů klasifikace   imunologie   MeSH
• prasata imunologie   MeSH
• receptory komplementu 3d imunologie   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH