19 results on '"Skjolaas, K."'
Search Results
2. Oral inoculation with Salmonella enterica serovar Typhimurium or Choleraesuis promotes divergent responses in the somatotropic growth axis of swine1
- Author
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Davis, B. L., primary, Fraser, J. N., additional, Burkey, T. E., additional, Skjolaas, K. A., additional, Dritz, S. S., additional, Johnson, B. J., additional, and Minton, J. E., additional
- Published
- 2010
- Full Text
- View/download PDF
3. BOARD-INVITED REVIEW: Porcine mucosal immunity of the gastrointestinal tract1
- Author
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Burkey, T. E., primary, Skjolaas, K. A., additional, and Minton, J. E., additional
- Published
- 2009
- Full Text
- View/download PDF
4. Effects of feeding Salmonella enterica serovar Typhimurium or serovar Choleraesuis on growth performance and circulating insulin-like growth factor-I, tumor necrosis factor-α, and interleukin-1β in weaned pigs1
- Author
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Fraser, J. N., primary, Davis, B. L., additional, Skjolaas, K. A., additional, Burkey, T. E., additional, Dritz, S. S., additional, Johnson, B. J., additional, and Minton, J. E., additional
- Published
- 2007
- Full Text
- View/download PDF
5. Growth performance of nursery pigs fed biosaf in combination with in-feed antimicrobials
- Author
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Hildabrand, B M, primary, Burkey, T E, additional, Skjolaas, K A, additional, Johnson, B J, additional, Minton, J Ernest, additional, and Dritz, Steven S, additional
- Published
- 2006
- Full Text
- View/download PDF
6. Effect of Quillaja saponaria extract on weanling pig growth performance and immune function during acute enteric disease challenge. (Nonruminant Nutrition)
- Author
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Turner, J.L., Dritz, S.S., Werner, J.R., Hill, C.M., Skjolaas, K., Herkelman, K., and Minton, J.E.
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Swine -- Growth ,Enterobacteriaceae -- Physiological aspects ,Immunological tolerance -- Research ,Zoology and wildlife conservation - Abstract
Quillaja saponaria (QS) extract has been in vaccine adjuvant for three Decades. The saponin fraction of the extract can inhibit E. coli growth. We investigated the effect of dietary QS extract (without added antibiotics) on growth and immune function of nursery pigs challenged with Salmonella typhimurium (STC). A total of 96 pigs (8.6 kg) were assigned to eight treatments in a 2 x 4 factorial with main effects of disease challenge (control vs. STC) and dietary treatment (0, 125, 250, or 500 mg/kg added QS). ADG, ADFI, and G/F were determined on d 7, 14, 21, and 28 of the trial. On d 14, 48 pigs were infected orally with S. typhimurium. Daily feed intake (DFI) and rectal temperature (RT) were monitored for 7 d following STC. On d 0, 7, and 14 after STC, serum was analyzed for haptoglobin (HAPT) and [[beta].sub.1]-acid glycoprotein (AGP). On d 0, 2, 4, and 6 post-challenge, serum IGF-I levels were determined. There were no differences (P>.05) in ADG, ADFI, G/F, DFI, RT, IGF-I, HAPT, or AGP between QS treatments. A STC by time interaction (P Key Words: Swine disease, IGF-I, Acute phase response
- Published
- 2001
7. Effect of seaweed extract on weanling pig growth performance and immune function during acute enteric disease challenge. (Nonruminant Nutrition)
- Author
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Turner, J.L., Dritz, S.S., Werner, J.R., Hill, C.M., Skjolaas, K., and Minton, J.E.
- Subjects
Swine -- Growth ,Immunological tolerance -- Research ,Marine algae as food -- Physiological aspects ,Zoology and wildlife conservation - Abstract
Extracts of seaweed plants have been shown to have antitumor effects in rodents. More recently the extract of the seaweed Ascophyllum nodosum (ANOD) has shown beneficial effects on fescue toxicosis in beef cattle. We investigated the effect of dietary ANOD (without added antibiotics) on growth and immune function of nursery pigs in the presence of Salmonella typhimurium challenge (STC). Pigs (6.8 kg) were blocked by weight and assigned randomly to one of eight treatments (two pigs per pen; six pens per treatment) in a 2 x 4 factorial with main effects of disease challenge (control vs. STC) and dietary treatment (0, 0.5. 1.0, or 2.0% added ANOD). ADG, ADFI, and G/F were determined on d 7, 14, 21, and 28 of the trial. On d 14, 48 pigs were infected orally with S. typhimurium. During the 7 d following STC, daily feed intake (DFI) and rectal temperature (RT) were monitored. On d 0, 7, and 14 with respect to STC, serum was analyzed for haptoglobin (HAPT) and [[beta].sub.1]-acid glycoprotein (AGP). There were no differences (P>.05) in ADG, ADFI, G/F, DFI, RT, HAPT, or AGP between ANOD treatments. A STC by time interaction (P Key Words: Swine disease, Seaweed, Immunity
- Published
- 2001
8. Effects of a seaweed extract on weanling pig growth performance and immune function during an acute enteric disease challenge
- Author
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Turner, J L, primary, Werner, J R, additional, Hill, C M, additional, Skjolaas, K, additional, Hogge, S, additional, Minton, J Ernest, additional, and Dritz, Steven S, additional
- Published
- 2000
- Full Text
- View/download PDF
9. Effects of a quillaja saponaria extract on weanling pig growth performance and immune function during an acute enteric disease challenge
- Author
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Turner, J L, primary, Werner, J R, additional, Hill, C M, additional, Skjolaas, K, additional, Hogge, S, additional, Herkleman, K, additional, Minton, J Ernest, additional, and Dritz, Steven S, additional
- Published
- 2000
- Full Text
- View/download PDF
10. Oral inoculation with Salmonella entericaserovar Typhimurium or Choleraesuis promotes divergent responses in the somatotropic growth axis of swine1
- Author
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Davis, B. L., Fraser, J. N., Burkey, T. E., Skjolaas, K. A., Dritz, S. S., Johnson, B. J., and Minton, J. E.
- Abstract
Enteric disease and immune challenge are processes that have detrimental effects on the growth performance of young swine. The current study tested the hypothesis that salmonella-induced enteric disease would perturb the endocrine growth axis in a serovar-dependent fashion. Specifically, we evaluated the effects of Salmonella entericaserovar Typhimurium (Typhimurium) and serovar Choleraesuis (Choleraesuis) on critical regulatory components of growth in young swine. Weaned pigs were housed 2 per pen with ad libitum access to feed and water in a 14-d experiment. Pigs were then repeatedly fed 108cfu of either Choleraesuis or Typhimurium in dough balls, with control pigs receiving dough without bacteria. Bacteria were refed twice weekly. Rectal temperatures were monitored daily from d 0 to 7 and ADFI was measured through d 14. Pigs were weighed and samples of serum were obtained for circulating IGF-I on d 0, 7, and 14. At the conclusion of the study, samples of semitendinosus muscle and liver were obtained and subsequently assayed for IGF-I, IGFBP-3, and IGFBP-5 mRNA. Rectal temperatures were elevated in pigs given Choleraesuis from d 2 through 7 (P< 0.05) when compared with control pigs and pigs fed Typhimurium. Pigs receiving Choleraesuis had a substantially decreased feed intake on d 2, 3, 4, 7, 8, 9, and 10 (P< 0.01), with a trend for a reduction on d 5 (P= 0.08), and they experienced an approximately 25% reduction in BW compared with control pigs and pigs given Typhimurium by the conclusion of the study. Pigs given Choleraesuis also experienced marked reductions in circulating IGF-I on d 7 (P< 0.01 vs. control and Typhimurium), with reductions of lesser magnitude on d 14 (P= 0.07 vs. control and P< 0.05 vs. Typhimurium). Inoculation tended to affect liver IGFBP-3 mRNA (P= 0.08), for which expression tended to be elevated in pigs given Typhimurium and Choleraesuis. In contrast, IGFBP-3 mRNA relative abundance was increased (P< 0.03) in pigs given Typhimurium compared with control pigs. Muscle IGF-I mRNA was reduced in pigs given Choleraesuis compared with control pigs and pigs given Typhimurium (P< 0.05). Treatment tended to affect muscle IGFBP-3 mRNA (P= 0.10). Oral inoculation of growing pigs with Choleraesuis disrupted feed intake and BW gain, and this was accompanied by decreases in circulating IGF-I and reduced muscle expression of mRNA for IGF-I and IGFBP-3.
- Published
- 2010
- Full Text
- View/download PDF
11. Effects of feeding Salmonella entericaserovar Typhimurium or serovar Choleraesuis on growth performance and circulating insulin-like growth factor-I, tumor necrosis factor-α, and interleukin-1β in weaned pigs1
- Author
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Fraser, J. N., Davis, B. L., Skjolaas, K. A., Burkey, T. E., Dritz, S. S., Johnson, B. J., and Minton, J. E.
- Abstract
The most common Salmonellaserovars causing clinical disease in pigs are Salmonella entericaserovars Typhimurium (Typhimurium) and Choleraesuis. Given that the swine host-adapted serovar Choleraesuis has been reported to cause systemic disease, a different disease outcome from that of Typhimurium, our working hypothesis was that this serovar would likely engage systemic immune-inflammatory mechanisms, resulting in elevated systemic cytokine secretion. Forty-eight weaned pigs were blocked by BW and sex, and randomly allotted to 1 of 3 treatments in a 14-d study. Each treatment had 8 replicates (pens), with 2 pigs/pen. The treatments consisted of a negative control and pigs repeatedly fed 108cfu of Typhimurium or Choleraesuis. On d 0, the pigs were fed Choleraesuis or Typhimurium in dough balls, and the bacteria were refed twice weekly throughout the experiment. Control pigs received dough balls without bacteria. All pigs were housed in temperature-controlled rooms under constant lighting and were fed a standard corn-soybean meal-based nursery diet. Pig BW and feed disappearance were used to determine ADG, ADFI, and G:F. Rectal temperatures were obtained daily from 1 pig/pen beginning 2 d before the first bacterial feeding through d 7 using rapid-response digital thermometers. Serum was collected on d 0, 7, and 14 from a single pig/pen for analysis of IGF-I, tumor necrosis factor-α , and IL-1β. There was no change in the rectal temperature of the control or the Typhimurium-challenged pigs (compared with d 0) or when comparing Typhimurium-challenged pigs with control animals. In contrast, pigs fed Choleraesuis had increased rectal temperatures beginning on d 2 and continuing through d 7 (P< 0.05), with the greatest elevation on d 3 (P< 0.001) compared with the control pigs. Average daily gain and ADFI of pigs challenged with Typhimurium did not differ from those of the control animals. Pigs fed Choleraesuis had a 25% reduction in ADG (P< 0.0001) and ADFI (P< 0.002) compared with the control pigs. On d 7, pigs fed Choleraesuis had reduced serum IGF-I compared with control (P< 0.01) or Typhimurium-challenged pigs (P= 0.01). Bacterial feeding did not affect serum tumor necrosis factor-α or IL-1β compared with control pigs at any time throughout the experiment. We conclude that repeated exposure of weaned pigs to Choleraesuis reduced growth performance in the absence of changes in systemic inflammatory cytokines.
- Published
- 2007
- Full Text
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12. Effects of dietary fish oil and flaxseed on plasma fatty acid composition and immune response in yearling horses.
- Author
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Vineyard, K. R., Warren, L. K., Skjolaas, K. A., Minton, J. E., and Kivipelto, J.
- Subjects
FISH oils ,LINSEED oil ,FATTY acids ,OMEGA-3 fatty acids ,HORSES ,IMMUNE response ,CONCANAVALIN A - Abstract
To determine the effects of different sources of dietary n-3 fatty acids on plasma FA composition and immune response in yearling horses, 18 Quarter Horse yearlings were randomly assigned to one of three treatments: encapsulated fish oil (FISH, n=6), milled flaxseed (FLAX, n=6), or no supplementation (CON, n=6). FISH contained 15 g eicosapentaenoic acid (EPA) and 12.5 g docosahexaenoic acid (DHA) and FLAX contained 61 g ∝-linolenic acid (ALA) per 100 g fat. Horses had free access to bahiagrass pasture and were individually fed a grain mix concentrate at 1.5% BW/d. FISH and FLAX were mixed into the concentrate in amounts to provide 6 g total n-3/100 kg BW. Horses were fed their respective treatments for 70 d. Blood samples were obtained at 0, 35 and 70 d for determination of plasma FA and isolation of peripheral blood mononuclear cells (PBMC). PBMC were stimulated with Concanavalin A and phytohemagglutinin (PHA) for determination of lymphocyte proliferation (LP). PBMC collected on d 70 were also challenged with lipopolysaccharide (LPS) to determine PGE2 production. On d 70, horses were injected intradermally with PHA, and skin thickness and area of swelling were evaluated over a 48 h period to assess in vivo inflammatory response. Treatment did not affect body weight gain (mean±SE, 41.8±1.9 kg). Horses fed FISH had higher (P<0.05) plasma EPA, DHA and total n-3 and lower (P<0.05) plasma linoleic acid, ALA and n-6:n-3 FA ratio than FLAX and CON. PBMC positively responded to mitogen stimulation and PGE2 increased in response to LPS, but treatment did not affect LP or PGE2 production. Across treatments, peak increase in skin thickness was observed between 4 h and 6 h after PHA injection. At 4 h post injection, FISH and FLAX had a greater increase in skin thickness than CON (P<0.05) and FISH had a larger area of swelling than CON at 4 h and 12 h (P<0.05). Although fed to supply a similar level of n-3 FA, FISH had a greater impact on plasma n-3 FA and n-6:n-3 ratio than FLAX. However, both FISH and FLAX demonstrated a more pronounced early inflammatory response to PHA injection than unsupplemented horses. [ABSTRACT FROM AUTHOR]
- Published
- 2006
13. Expression of porcine Toll-like receptor 2, 4 and 9 gene transcripts in the presence of lipopolysaccharide and Salmonella enterica serovars Typhimurium and Choleraesuis.
- Author
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Burkey TE, Skjolaas KA, Dritz SS, and Minton JE
- Subjects
- Animals, Animals, Newborn, Cell Line, Epithelial Cells, Gastrointestinal Diseases genetics, Gastrointestinal Diseases immunology, Gastrointestinal Diseases microbiology, Gene Expression Profiling methods, Ileum immunology, Ileum microbiology, Lipopolysaccharides pharmacology, Male, RNA, Messenger genetics, Reverse Transcriptase Polymerase Chain Reaction veterinary, Salmonella Infections, Animal immunology, Salmonella Infections, Animal microbiology, Swine, Swine Diseases immunology, Swine Diseases microbiology, Toll-Like Receptors biosynthesis, Toll-Like Receptors immunology, Gastrointestinal Diseases veterinary, Gene Expression Profiling veterinary, RNA, Messenger biosynthesis, Salmonella Infections, Animal genetics, Salmonella typhimurium immunology, Swine Diseases genetics, Toll-Like Receptors genetics
- Abstract
Salmonella enterica serovar Typhimurium (ST) and Choleraesuis (SC) are among the most frequently isolated salmonellae serovars causing enteric disease in swine. Enteric disease in young pigs is of major concern in modern production systems due to the negative implications on animal health, food safety and economic return. Epithelial cells express Toll-like receptors (TLR) that recognize conserved microbial structures and act as mediators of innate and adaptive immune responses. However, little is known about the expression of TLR gene transcripts in swine. The objective of the current study was to characterize the relative abundance of porcine TLR2, 4 and 9 gene transcripts in vitro in a porcine jejunal epithelial cell line (IPEC-J2) and in porcine mononuclear phagocytes (pMP) in the presence of ST or SC, as well as in vivo in the distal ileum of pigs orally challenged with ST. Our results indicate that TLR2, 4 and 9 are constitutively expressed in vitro in IPEC-J2 cells and pMP and in vivo in the distal ileum. Additionally, transient modulation of porcine TLR was observed in vitro and in vivo in the presence of ST and SC. Further investigation is warranted to determine the effects of ST and SC on functional TLR.
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- 2009
- Full Text
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14. Effects of feeding Salmonella enterica serovar Typhimurium or serovar Choleraesuis on growth performance and circulating insulin-like growth factor-I, tumor necrosis factor-alpha, and interleukin-1beta in weaned pigs.
- Author
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Fraser JN, Davis BL, Skjolaas KA, Burkey TE, Dritz SS, Johnson BJ, and Minton JE
- Subjects
- Animal Feed, Animals, Diet veterinary, Female, Male, Salmonella genetics, Salmonella Infections, Animal metabolism, Species Specificity, Swine growth & development, Swine microbiology, Swine Diseases metabolism, Swine Diseases microbiology, Weight Gain, Insulin-Like Growth Factor I metabolism, Interleukin-1beta metabolism, Salmonella physiology, Salmonella Infections, Animal microbiology, Swine metabolism, Tumor Necrosis Factor-alpha metabolism
- Abstract
The most common Salmonella serovars causing clinical disease in pigs are Salmonella enterica serovars Typhimurium (Typhimurium) and Choleraesuis. Given that the swine host-adapted serovar Choleraesuis has been reported to cause systemic disease, a different disease outcome from that of Typhimurium, our working hypothesis was that this serovar would likely engage systemic immune-inflammatory mechanisms, resulting in elevated systemic cytokine secretion. Forty-eight weaned pigs were blocked by BW and sex, and randomly allotted to 1 of 3 treatments in a 14-d study. Each treatment had 8 replicates (pens), with 2 pigs/pen. The treatments consisted of a negative control and pigs repeatedly fed 10(8) cfu of Typhimurium or Choleraesuis. On d 0, the pigs were fed Choleraesuis or Typhimurium in dough balls, and the bacteria were refed twice weekly throughout the experiment. Control pigs received dough balls without bacteria. All pigs were housed in temperature-controlled rooms under constant lighting and were fed a standard corn-soybean meal-based nursery diet. Pig BW and feed disappearance were used to determine ADG, ADFI, and G:F. Rectal temperatures were obtained daily from 1 pig/pen beginning 2 d before the first bacterial feeding through d 7 using rapid-response digital thermometers. Serum was collected on d 0, 7, and 14 from a single pig/pen for analysis of IGF-I, tumor necrosis factor-alpha , and IL-1beta. There was no change in the rectal temperature of the control or the Typhimurium-challenged pigs (compared with d 0) or when comparing Typhimurium-challenged pigs with control animals. In contrast, pigs fed Choleraesuis had increased rectal temperatures beginning on d 2 and continuing through d 7 (P < 0.05), with the greatest elevation on d 3 (P < 0.001) compared with the control pigs. Average daily gain and ADFI of pigs challenged with Typhimurium did not differ from those of the control animals. Pigs fed Choleraesuis had a 25% reduction in ADG (P < 0.0001) and ADFI (P < 0.002) compared with the control pigs. On d 7, pigs fed Choleraesuis had reduced serum IGF-I compared with control (P < 0.01) or Typhimurium-challenged pigs (P = 0.01). Bacterial feeding did not affect serum tumor necrosis factor-alpha or IL-1beta compared with control pigs at any time throughout the experiment. We conclude that repeated exposure of weaned pigs to Choleraesuis reduced growth performance in the absence of changes in systemic inflammatory cytokines.
- Published
- 2007
- Full Text
- View/download PDF
15. Effects of Salmonella enterica serovar Typhimurium, or serovar Choleraesuis, Lactobacillus reuteri and Bacillus licheniformis on chemokine and cytokine expression in the swine jejunal epithelial cell line, IPEC-J2.
- Author
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Skjolaas KA, Burkey TE, Dritz SS, and Minton JE
- Subjects
- Animals, Chemokines biosynthesis, Chemokines immunology, Chemokines, CC genetics, Chemokines, CC immunology, Cytokines immunology, Enzyme-Linked Immunosorbent Assay veterinary, Epithelial Cells immunology, Epithelial Cells metabolism, Epithelial Cells microbiology, Interleukin-8 genetics, Interleukin-8 immunology, Jejunum cytology, Jejunum microbiology, RNA, Bacterial chemistry, RNA, Bacterial genetics, Reverse Transcriptase Polymerase Chain Reaction veterinary, Tumor Necrosis Factor-alpha genetics, Tumor Necrosis Factor-alpha immunology, Bacillus immunology, Cytokines biosynthesis, Jejunum immunology, Limosilactobacillus reuteri immunology, Salmonella arizonae immunology, Salmonella typhimurium immunology, Swine immunology
- Abstract
Direct-fed microbials, including Lactobacillus and Bacillus spp., are potential replacements for low dose in-feed antibiotics for swine and other livestock. To understand the function of these microbes in the gut, the current study used pig jejunal epithelial cells (IPEC-J2) to evaluate how Lactobacillus reuteri (LR) and Bacillus licheniformis (BL) differed from Salmonella enterica serovars Typhimurium (ST) or Choleraesuis (SC) in their ability to regulate, stimulate, or modify the proinflammatory mediators, interleukin 8 (IL8), CC chemokine 20 (CCL20), and tumor necrosis factor-alpha (TNFalpha). To optimize the positive control to drive IL8 secretion by IPEC-J2 cells, cells were treated apically with various concentrations of ST (versus control (CTL)) for 1h, followed by a wash. Media containing gentamicin was added and collected at 6h post-treatment. Compared to CTL, 10(8) ST produced maximal IL8 secretion in both the apical and basolateral directions, with significant basolateral polarization (P<0.0001). We next evaluated the time course of IL8 secretion, and IL8, CCL20, and TNFalpha mRNA expression by IPEC-J2 cells treated apically with 10(8) ST, SC, LR, and BL versus CTL. Media and RNA were collected at 1.5, 3.0, and 6.0 h post treatment. Only ST stimulated an increase in IL8 secretion at any time point, with increases in IL8 mRNA at both 3 and 6h (P<0.05). However, BL increased IL8 mRNA at 1.5h (P<0.0001). Neither LR nor SC affected IL8 mRNA expression. CCL20 mRNA was strongly upregulated by ST (P<0.05) and BL (1.5 and 3.0 h; P<0.05), but not LR or SC. Only ST increased TNFalpha mRNA relative to CTL (P<0.05). Two experiments were conducted to determine if pre-exposure of IPEC-J2 cells to LR or BL modified ST induced IL8 secretion. Confluent cells were treated apically overnight with various levels of LR or BL (in separate experiments) followed by ST challenge. Media were collected at 4 (LR experiment) or 5h (BL experiment) post ST. In the LR study, IL8 secretion was increased by ST as compared to CTL (P<0.0001), reduced by LR (P<0.05), and LR+ST co-treatments failed to alter ST stimulated secretion. In the BL experiment, secretion of IL8 was increased by ST (P<0.0001), but blunted basolaterally in BL+ST co-treated wells. The data demonstrate that IPEC-J2 cells increase IL8 secretion in response to ST, and IL8 mRNA in response to ST and BL, but not LR. Furthermore, ST stimulated secretion of IL8 is inhibited basolaterally in the presence of BL.
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- 2007
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16. Expression of Toll-like receptors, interleukin 8, macrophage migration inhibitory factor, and osteopontin in tissues from pigs challenged with Salmonella enterica serovar Typhimurium or serovar Choleraesuis.
- Author
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Burkey TE, Skjolaas KA, Dritz SS, and Minton JE
- Subjects
- Animals, Chemokines, CC genetics, Chemokines, CC immunology, Interleukin-8 biosynthesis, Interleukin-8 genetics, Interleukin-8 immunology, Intestinal Diseases immunology, Intestinal Diseases microbiology, Luminescent Proteins genetics, Macrophage Migration-Inhibitory Factors genetics, Macrophage Migration-Inhibitory Factors immunology, Osteopontin genetics, Osteopontin immunology, RNA, Messenger biosynthesis, RNA, Messenger genetics, Random Allocation, Reverse Transcriptase Polymerase Chain Reaction veterinary, Salmonella Infections, Animal microbiology, Salmonella typhimurium genetics, Swine, Swine Diseases immunology, Toll-Like Receptors genetics, Toll-Like Receptors immunology, Transformation, Genetic, Intestinal Diseases veterinary, Macrophage Migration-Inhibitory Factors biosynthesis, Osteopontin biosynthesis, Salmonella Infections, Animal immunology, Salmonella arizonae immunology, Salmonella typhimurium immunology, Swine Diseases microbiology, Toll-Like Receptors biosynthesis
- Abstract
Two serovars of Salmonella enterica, namely serovar Typhimurium (ST) and serovar Choleraesuis (SC) account for the vast majority of clinical cases of swine salmonellosis worldwide. These serovars are thought to be transmitted among pigs in production settings mainly through fecal-oral routes. Yet, few studies have evaluated effects of these serovars on expression of innate immune targets when presented to pigs via repeated oral dosing in an attempt to model transmission in production settings. Thus, a primary objective of the current experiments was to evaluate expression of Toll-like receptors (TLR) and selected chemoattractive mediators (interleukin 8, IL8; macrophage migration inhibitory factor, MIF; osteopontin, OPN) in tissues from pigs exposed to ST or SC that had been transformed with kanamycin resistance and green (STG) or red (SCR) fluorescent protein to facilitate isolation from pen fecal samples. In vitro studies confirmed that STG and SCR largely (though not completely) retained their ability to upregulate IL8 and CC chemokine ligand 20 (CCL20) in cultured swine jejunal epithelial cells. Transformed bacteria were then fed to pigs in an in vivo study to determine tissue specific effects on mRNA relative expression. Pigs were fed cookie dough inoculated with bacteria on days 0, 3, 7, and 10 with 10(8)CFU STG (n=8) or SCR (n=8), while control (CTL) pigs (n=8) received dough without bacteria. Animals were sacrificed 14 days from the initial bacterial challenge and samples of tonsil, jejunum, ileum, colon, mesenteric lymph node (MLN), spleen, and liver were removed for subsequent RNA isolation. Expression of mRNA in tissues was determined using real-time quantitative PCR and expressed relative to 18S rRNA. Within CTL pigs, when expressed relative to the content in liver, mRNA for all targets demonstrated substantial tissue effects (P<0.001 for all TLR; MIF, and OPN; P<0.05 for IL8). Feeding STG and SCR resulted in significant (P
- Published
- 2007
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17. Effects of Salmonella enterica serovars Typhimurium (ST) and Choleraesuis (SC) on chemokine and cytokine expression in swine ileum and jejunal epithelial cells.
- Author
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Skjolaas KA, Burkey TE, Dritz SS, and Minton JE
- Subjects
- Animals, Chemokines, CC genetics, Chemokines, CC immunology, Cytokines genetics, Cytokines immunology, Epithelial Cells immunology, Epithelial Cells microbiology, Gastrointestinal Diseases immunology, Gastrointestinal Diseases microbiology, Ileum immunology, Ileum microbiology, Jejunum immunology, Jejunum microbiology, Lipopolysaccharides immunology, Lipopolysaccharides pharmacology, Macrophage Migration-Inhibitory Factors genetics, Macrophage Migration-Inhibitory Factors immunology, Male, RNA, Messenger biosynthesis, RNA, Messenger genetics, Reverse Transcriptase Polymerase Chain Reaction veterinary, Salmonella Infections, Animal immunology, Swine, Swine Diseases immunology, Chemokines, CC biosynthesis, Cytokines biosynthesis, Gastrointestinal Diseases veterinary, Salmonella Infections, Animal microbiology, Salmonella arizonae immunology, Salmonella typhimurium immunology, Swine Diseases microbiology
- Abstract
The gastrointestinal epithelium represents a barrier to potentially invasive enteric pathogens, maintains a role in innate immune surveillance, and is a source of both chemokine and cytokine chemotactic mediators in response to bacterial invasion. In the current study, we evaluated cytokine and chemokine mediators known to regulate movement of macrophages (macrophage migration inhibitory factor; MIF), neutrophils (IL8), dendritic cells (CCL20), and epithelial remodeling (osteopontin; OPN) in response to invasive swine enteropathogens Salmonella enterica serovar Typhimurium (ST) or Choleraesuis (SC). For the in vivo experiment, weaned pigs served as uninfected controls (0 h) or were given 3 x 10(9) CFU ST orally. Pigs were sacrificed at 8, 24, 48, and 144 h after inoculation and total RNA was extracted from defined segments of proximal (PI) and distal (DI) ileum. Relative expression of MIF and OPN were not affected by ST. IL8 expression was increased numerically (P = 0.17 for the interaction term) at 24 and 144 h in the PI and these increases accounted for greater expression in the PI relative to the DI (P < 0.05). Relative expression of CCL20 was increased at 24 h after ST (P < 0.05). Next, we evaluated the time course of MIF, IL8, CCL20, and OPN mRNA expression induced by application of lipopolysaccharide (LPS), ST or SC in vitro using pig jejunal epithelial cells (IPEC-J2). Cells were grown to confluency on permeable membranes, and treated apically with LPS (10 ng/mL), ST or SC (10(8)/well). After 1 h, cells were washed to remove LPS or extracellular bacteria, and media containing gentamicin was added to kill remaining extracellular bacteria. Media and RNA were collected at 1.5, 3, and 6 h after treatment. MIF mRNA was not affected by LPS or bacterial treatment. Similarly, IL8 expression was not affected by LPS, but was increased by ST and SC relative to controls at 1.5 and 3 h post exposure (P < 0.05 for all comparisons). Treatment with SC increased CCL20 mRNA relative to controls at 3 h (P < 0.05), while ST increased CCL20 at 1.5, 3, and 6h with maximal expression at 6 h (P < 0.05 for all comparisons). ST and SC increased polarized IL8 secretion. Our data demonstrate that invasive bacterial pathogens in the pig gastrointestinal tract trigger upregulation of selected cytokine and chemokine mediators, but serovars of Salmonella elicited differing patterns of activation in vitro.
- Published
- 2006
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18. Does cortisol bias cytokine production in cultured porcine splenocytes to a Th2 phenotype?
- Author
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Skjolaas KA and Minton JE
- Subjects
- Animals, Cells, Cultured, Glucocorticoids pharmacology, Lymphocyte Activation, Macrophage Migration-Inhibitory Factors biosynthesis, Cytokines biosynthesis, Hydrocortisone biosynthesis, Spleen cytology, Swine immunology, Th1 Cells immunology, Th2 Cells immunology
- Abstract
Glucocorticoids are reported to bias the production of cytokines from a type 1 to a type 2 phenotype. However, this dogma has been advanced largely from studies utilizing potent glucocorticoid analogs, particularly dexamethasone (DEX). Although studies utilizing DEX certainly have clinical and pharmacological relevance, DEX is probably not the best glucocorticoid for studies designed to evaluate the interaction and regulation of endogenous corticosteroids with immune cells in vivo in the domestic pig. Functional measures of immune suppression suggest that the pig is relatively resistant to DEX. Furthermore, type II corticosteroid receptors exclusively bind DEX with high affinity, whereas type I receptors, the so-called mineralocorticoid receptors, have a higher affinity for cortisol. In addition, DEX is not bound by serum binding proteins as are endogenous corticosteroids. These issues prompted us to revisit glucocorticoid regulation of type 1 and type 2 cytokines in cultured pig splenocytes and to test the broad hypothesis that cortisol biases cytokine production in favor of a Th2 response. We evaluated interferon gamma (IFNgamma) (also interleukin 2 (IL-2) in one experiment) and interleukin 10 (IL-10) as representative Th1 and Th2 cytokines, respectively. Furthermore, we evaluated macrophage migration inhibitory factor (MIF) because it is reported to be an essential factor in T cell activation; it is also upregulated by glucocorticoids and reported to be a product of Th2 lymphocytes. In general, both IFNgamma and IL-10 were sensitive to cortisol inhibition early in culture. However, IFNgamma ultimately escaped cortisol inhibition, whereas IL-10 continued to be substantially suppressed by high physiological concentrations of cortisol. Similarly, MIF mRNA could be suppressed by cortisol, but only when cortisol was added to cultures after ConA (concanavalin A) stimulation of splenocytes. So, taken together, our studies do not support the hypothesis that cortisol favors a Th2 cytokine profile in cultured pig splenocytes.
- Published
- 2002
- Full Text
- View/download PDF
19. Glucocorticoid regulation of type 1 and type 2 cytokines in cultured porcine splenocytes.
- Author
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Skjolaas KA, Grieger DM, Hill CM, and Minton JE
- Subjects
- Animals, Blotting, Northern veterinary, Cells, Cultured, Dexamethasone pharmacology, Female, Glucocorticoids pharmacology, Interferon-gamma immunology, Interleukin-10 immunology, Interleukin-2 immunology, Macrophage Migration-Inhibitory Factors biosynthesis, Macrophage Migration-Inhibitory Factors genetics, Macrophage Migration-Inhibitory Factors immunology, Male, RNA, Messenger metabolism, Th1 Cells drug effects, Th1 Cells immunology, Th1 Cells metabolism, Th2 Cells drug effects, Th2 Cells immunology, Th2 Cells metabolism, Anti-Inflammatory Agents pharmacology, Hydrocortisone pharmacology, Interferon-gamma biosynthesis, Interleukin-10 biosynthesis, Interleukin-2 biosynthesis, Swine immunology
- Abstract
Glucocorticoids are reported to bias cytokines to a Th2 phenotype. However, this dogma has been advanced largely from studies utilizing potent glucocorticoid analogs. The current study was conducted to revisit the issue of glucocorticoid modulation of Th1/Th2 cytokine production and evaluate migration inhibitory factor (MIF) mRNA expression in cultured pig splenocytes treated with physiologically relevant concentrations of cortisol (CORT). Dexamethasone (DEX) was included for comparison. In Experiment 1, DEX, at 150 and 300 nM, suppressed concanavalin (ConA)-stimulated IFNgamma at both 12 and 24 h in culture, and IL-10 at 24h (P<0.05). Both 150 and 300 nM CORT suppressed IL-10 at 24 h (P<0.05), but neither concentration affected IFNgamma at 24 h. In Experiment 2, cells were cultured with a broader range of CORT for 48 h following ConA. Parallel cultures with identical treatments also were conducted in separate plates for evaluation of glucocorticoid regulation of MIF mRNA. IFNgamma was reduced by 300 nM DEX at 12, 24, and 48 h (P<0.05), whereas 150 and 300 nM CORT blunted IFNgamma at 24 h (P<0.05), but not 48 h. ConA increased IL-2 (P<0.01), but none of the steroid treatments affected IL-2. At both 12 and 24 h, IL-10 was reduced by 300 nM DEX and by 150 and 300 nM CORT (P<0.05). ConA increased relative abundance of MIF mRNA (P<0.001), but no steroid treatment affected MIF mRNA. In Experiment 3, steroid additions were delayed by 24 h after ConA, and cytokine concentrations evaluated 48 h later. Again, separate cultures were used for determination of effect of treatments on MIF mRNA. None of the steroid treatments affected IFNgamma, but 300 nM DEX reduced IL-10 (P<0.05). All of the CORT treatments (75-300 nM) reduced MIF mRNA (P<0.05), whereas DEX did not affect MIF mRNA in this experiment. The current experiments suggest that both DEX and high physiological concentrations of CORT can suppress both type 1 and type 2-like cytokines in cultured pig splenocytes. But, IL-10 was generally more sensitive to CORT suppression with increased time in culture than was IFNgamma. In addition, MIF mRNA could be suppressed by delayed addition of CORT to porcine splenocytes. Taken together, the data do not support the hypothesis that CORT directs the cytokine milieu toward a type 2 bias in cultured pig splenocytes.
- Published
- 2002
- Full Text
- View/download PDF
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