Probiotic fermentation of Ganoderma lucidum fruiting body extracts promoted its immunostimulatory activity in mice with dexamethasone-induced immunosuppression.

Ganoderma lucidum is a legendary traditional Chinese medicine with various bioactivities. This study was conducted (a) to explore the in vitro fermentation of the water extracts of G. lucidum fruiting body with Lactobacillus acidophilus and Bifidobacterium breve and (b) to investigate the effect of fermentation broth (GLFB) on dexamethasone (DEX)-induced immunosuppressed mice. Our results demonstrated that probiotic fermentation of G. lucidum fruiting body extracts underwent structural changing of major ganoderic acid components, such as ganoderic acid A (GA) into GC2, and this fermentation process involves changing of several metabolic pathways in the probiotic strains. GLFB could significantly improve the immunity, intestinal integrity, and gut microbiota dysbiosis in DEX-treated mice, and the immunostimulatory activity of GLFB was found closely related to its direct regulation on the expansion of CD4+ T cells in Peyer's patches of mice. These data implied that probiotic fermentation of G. lucidum fruiting body extracts promoted its immunostimulatory activity via biotransformation of components such as GA. This research provides a theoretical support for the development and application of G. lucidum fermentation by probiotics.

Characteristics and intestinal immunomodulating activities of water-soluble pectic polysaccharides from Chenpi with different storage periods.

BACKGROUND: The traditional view considers that Chenpi (dried citrus peel) stored over the long-term has better health efficacies compared to fresh Chenpi, although the detailed mechanism responsible for this remains obscure. RESULTS: The three water-soluble pectic polysaccharides (CPP1, CPP5 and CPP10) were obtained from 1-, 5- and 10-year Chenpi, respectively, and their physicochemical characteristics and intestinal immunomodulating activities were investigated and compared. The results obtained showed that CPP5 and CPP10 demonstrated a lower dynamic viscosity and degree of methylesterification, as well as a higher molecular heterogeneity, compared to CPP1. Monosaccharide composition analysis indicated that CPP1 was composed of arabinose, galacturonic acid and galactose, and a small amount of rhamnose; however, CPP5 and CPP10 consisted of arabinose, galacturonic acid, galactose, glucose and xylose, and a small amount of rhamnose. With the extension of storage period of Chenpi, the content of soluble conjugate phenolic acids increased in the pectic polysaccharide. Furthermore, it was confirmed that the pectic polysaccharides extracted from the 5-year and 10-year Chenpi could significantly enhance the proliferation of bone marrow cells via activating the Peyer's patch cells in vitro. CONCLUSION: The present study demonstrates the differences in the pectic polysaccharides from Chenpi with different storage periods and also confirms that the pectic polysaccharides extracted from Chenpi stored over the long-term had more significant intestinal activities compared to that obtained from the fresh Chenpi. This phenomenon might partly explain why the Chenpi stored over the long-term has better healthcare effects. © 2018 Society of Chemical Industry.

The common lavender (Lavandula angustifolia Mill.) pectic polysaccharides modulate phagocytic leukocytes and intestinal Peyer's patch cells.

Two pectic (chPS-L1, chPS-L2) and one polyphenolic (chPP-L) fractions were obtained from lavender flowers after boiling water extraction, exhaustive removing of alcohol-soluble molecules and SEC. chPS-L1 (52.4kDa) contains mainly low-acetylated and high-methoxylated homogalacturonans (HG), and smaller rhamnogalacturonan (RG) I backbone fragments rich in 1,3,5-branched arabinan and arabinogalactan (AG) II side chains. chPS-L2 (21.8kDa) contains predominantly similarly esterified HG, followed by RGI with AGII structures and RGII. The prevalence of catechin and epicatechin in chPP-L indicates that they form weak interactions with pectins. chPS-L1 and chPS-L2 enhanced ß2-integrin expression on neutrophils, inducing ROS generation and macrophage NO production. Both the effects on ß2-integrin and high complement fixation activity of chPS-L1 were proposed for its inhibitory action against PMA- and OZP-activated ROS formation. This, together with suppression of NO generation after co-stimulation with chPS-L1 and LPS, suggested anti-inflammatory activity of studied pectins. Lavender polysaccharides expressed intestinal Peyer's patch immunomodulating activity.

[Shengqifuzheng Injection promotes the recovery of B cells in gut-associated lymphoid tissues of mice treated with cyclophosphamide].

Objective To investigate the effect of Shengqifuzheng Injection (SQFZ) on the number recovery of B cells in gut-associated lymphoid tissues (GALTs) of mice receiving cyclophosphamide-based chemotherapy. Methods BALB/c mice were randomly divided into control group, cyclophosphamide (Cy) group and SQFZ group. Mice in Cy group and SQFZ group were injected intraperitoneally with Cy (100 mg/kg), while the control mice were injected with an equal volume of normal saline. Twenty-four hours later, mice in SQFZ group were administrated intragastricly with 1 mL SQFZ once daily for 10 consecutive days, and mice in the other groups were given the same volume of normal saline. Body mass of all the mice was measured every day. Mice were killed on day 10, and the indexes of spleen and thymus were measured. Cell cycles of bone marrow cells and the percentage of B cells in lymphocytes in mesenteric lymph node (MLN) and Peyer's patch (PP) were detected by flow cytometry. In vitro, after being treated with SQFZ, activity of lymphocytes was evaluzed by MTT assay; expression of CD86 on B cell surface was analyzed by flow cytometry; and B cell proliferation was tested by carboxyfluorescein succinimidyl ester (CFSE)-based lymphocyte proliferation assay. Results SQFZ alleviated the loss of body mass caused by Cy and promoted the recovery of thymus indexes, spleen indexes and B cell number in MLN and PP. But it did not alleviate the bone marrow suppression of mice in this condition. In vitro, SQFZ enhanced lymphocyte activity, and improved the activation and proliferation of B cells. Conclusion SQFZ could accelerate the recovery of B cells in GALTs of mice receiving chemotherapy and it might act by promoting B cell proliferation.

Intravenous administration of high-dose Paclitaxel reduces gut-associated lymphoid tissue cell number and respiratory immunoglobulin A concentrations in mice.

BACKGROUND: Chemotherapy remains a mainstay of treatment for cancer patients. However, anti-cancer drugs frequently cause a wide range of side effects, including leukopenia and gastrointestinal toxicity. These adverse effects can lead to treatment delays or necessitate temporary dose reductions. Although chemotherapy-related changes in gut morphology have been demonstrated, the influences of chemotherapeutic regimens on gut immunity are understood poorly. This study aimed to examine whether the anti-cancer drug paclitaxel (PTX) impairs gut immunity in mice. METHODS: Male ICR mice were randomized into three groups: Control, low-dose PTX (low PTX; 2 mg/kg), or high-dose PTX (high PTX; 4 mg/kg). A single intravenous dose was given. On day seven after the injection, lymphocytes from Peyer patches (PP), intraepithelial (IE) spaces, and the lamina propria (LP) were counted and analyzed by flow cytometry (CD4(+), CD8(+), αßTCR(+), γδTCR(+), B220(+)). Immunoglobulin A (IgA) concentrations were measured in small intestinal and respiratory tract washings. RESULTS: Total, CD4(+) and γδTCR(+) lymphocyte numbers in PPs were significantly lower in the high PTX than in the control group. The CD4(+) lymphocyte numbers in the IE spaces were significantly lower in both PTX groups than in the control group. Respiratory tract IgA concentrations were lower in the high PTX than in the control group. CONCLUSION: The present data suggest high-dose PTX impairs mucosal immunity, possibly rendering patients more vulnerable to infection. Careful dose selection and new therapies may be important for maintaining mucosal immunity during PTX chemotherapy.

Immunomodulating pectic polysaccharides from waste rose petals of Rosa damascena Mill.

A water-soluble polysaccharide (RP-1) was obtained from distilled rose petals of Rosa damascena Mill. as an attempt for valorization of the waste. RP-1 showed in vitro intestinal immune system modulating activity through Peyer's patch cells and IL-6 producing activity from macrophages. RP-1 lost most of its immunomodulating activity by degradation of the carbohydrate moiety with periodate. RP-1 was fractionated by anion-exchange and gel filtration chromatography and some of the fractions showed significant intestinal immune system modulating activity. The active fractions were suggested to be pectic polysaccharides and type II arabino-3,6-galactan from the component sugar analyses and the reactivity with Yariv antigen. When some active fractions were digested with endo α-d-(1→4)-polygalacturonase, highest molecular weight fragments which were considered as rhamnogalacturonan I, showed potent immunomodulating activities. To our knowledge, this is a first report which explores the possibility for utilization of waste rose petals as a source of immunomodulating pectic polysaccharides.

Augmentation of cellular immunity and protection against influenza virus infection by bovine late colostrum in mice.

OBJECTIVE: We investigated whether oral administration of skimmed and concentrated bovine late colostrum (SCBLC) activates the immune system and protects against influenza virus (Flu) infection. METHODS: Murine Peyer's patch (PP) cells (2.5 105) were cultured in 0.1 ml RPMI-1640 supplemented with SCBLC at a concentration of 0, 0.1 or 1.0 mg/ml. To determine the levels of IL-12 and IFN-, supernatants were collected on day 3. Mice were orally administered sterile saline solution (control group), or 400 g/g body weight (SCBLC 400 group) or 2,000 g/g body weight (SCBLC 2,000 group) of SCBLC for three weeks. These mice were measured for natural killer (NK) cells activity on PP cells, splenocytes and lung cells. Also, these mice in the control and SCBLC 2,000 groups were infected with Flu and were measured for the accumulated symptom rate. RESULTS: In PP cells cultured with SCBLC, the levels of IL-12 and IFN- were significantly increased in vitro. Oral administration of SCBLC to mice significantly increased NK cell activity of PP cells, splenocytes and lung cells. The accumulated symptom rate of the SCBLC 2,000 group was significantly lower than that of the control group in a mouse model of Flu infection. CONCLUSION: These results indicate that oral administration of SCBLC activates not only systemic cellular immunity but also local cellular immunity, such as in the respiratory tract, and that activation of cellular immunity is one of the mechanisms of amelioration of Flu infection.

Phenolic lipid ingredients from cashew nuts.

Five new phenolic lipids, 2-(8"Z-eicosenoyl)-6-(8'Z-pentadecenyl) salicylic acid (3), 2-(9"Z-hexadecenoyl)-6-(8'Z, 11'Z-pentadecadienyl) methyl salicylate (5), 2-(10"Z, 13"Z-nonadecadienoyl)-6-(8'Z, 11'Z-pentadecadienyl) salicylic acid (6), 2-(16"Z-pentacosenoyl)-6-(8'Z-pentadecenyl) salicylic acid (7) and 2-(9"Z-octadecenoyl)-6-(8'Z, 11'Z-pentadecadienyl) methyl salicylate (8), and three known compounds, cardols (1), anacardic acid (2) and cardanols (4), were isolated from the nuts of the cashew, Anacardium occidentale L. The structures were established on the basis of detailed MS and NMR spectroscopic analyses. Compound 1 highly enhanced both Th-1 (IL-2, IFN-γ) and Th-2 (IL-4, IL-5) cytokine production, and compounds 7 and 8 highly increased cytokine IL-2 and IFN-γ production in response to concanavalin A in cultured murine Peyer's patch cells ex vivo. The isolated compounds showed moderate inhibitory activities on cytochrome CYP3A4 enzyme.

Dietary ß-glucan regulates the levels of inflammatory factors, inflammatory cytokines, and immunoglobulins in interleukin-10 knockout mice.

ß-Glucan is known to have anti-inflammatory properties, and several studies have demonstrated the beneficial effects of dietary ß-glucan on inflammatory bowel disease (IBD). However, it is unknown how ß-glucan mediates its protective effects on IBD. Therefore, we used a well-established mouse model for IBD, interleukin (IL)-10(-/-) mice, to explore the protective effects of ß-glucan on IBD-like symptoms caused by IL-10 deficiency. The mice were divided into two groups: IL-10(-/-) and IL-10(-/-) + ß-glucan treatment groups. IL-10(-/-) mice treated with dietary ß-glucan exhibited less inflammation within the colon. The levels of immunoglobulins A and E were lower in the serum, spleen, mesenteric lymph nodes, and Peyer's patches in the IL-10(-/-) mice compared with the IL-10(-/-) + ß-glucan mice. Also, the expression of pro-inflammatory cytokines was lower in the IL-10(-/-) + ß-glucan mice compared with the IL-10(-/-) mice. Histological analysis also revealed that administration of dietary ß-glucan in IL-10(-/-) mice reduced colonic tissue damage. Finally, the expression of the pro-inflammatory cytokine tissue necrosis factor-α was significantly lower with dietary ß-glucan treatment in IL-10(-/-) mice. In conclusion, dietary ß-glucan reduces the inflammation associated with IBD caused by IL-10 deficiency.

Effect on the epigallocatechin gallate/epigallocatechin ratio in a green tea (Camellia sinensis L.) extract of different extraction temperatures and its effect on IgA production in mice.

We found that the epigallocatechin gallate (EGCG)/epigallocatechin (EGC) ratio in a green tea (Camellia sinensis L.) extract was affected by the extraction temperature. The EGCG/EGC ratio in the 4 °C extract was around 1:3-4, whereas in the 100 °C extract, it was around 1:0.7. Oral administration of the mixture with a high EGC ratio (1:2-3 = EGCG/EGC) resulted in greater IgA production by murine Peyer's patch cells.

Effects of enteral nutrition supplemented with glutamine on intestinal mucosal immunity in burned mice.

OBJECTIVE: The aim of this study was to investigate the effects of enteral nutrition (EN) supplemented with glutamine (GLN) on Peyer's patches and intestinal immunoglobulin A (IgA) response in burned mice. METHODS: Thirty-four mice were randomly assigned to a normal control group (n = 10), an EN group (n = 12), and an EN supplemented with GLN (EN + GLN) group (n = 12) and mice in the EN and EN + GLN groups received a 20% total body surface area, full-thickness scald burn on the back. Then the burned mice were fed with conventional EN or EN + GLN for 7 d. There was isonitrogenous and isocaloric intake in the EN and EN + GLN groups. On day 7 after injury, entire intestines were collected and intestinal IgA levels, total lymphocyte yield, lymphocyte subpopulations, and total apoptotic ratio in Peyer's patches were analyzed. RESULTS: Total lymphocyte yield, numbers of lymphocyte subpopulations, and intestinal IgA levels in the EN + GLN group were significantly higher than those in the EN group (P < 0.05). The total apoptotic ratio in Peyer's patches was markedly decreased in the EN + GLN group compared with that in the EN group (P < 0.05). CONCLUSION: The results indicated that EN supplemented with GLN is superior to conventional EN with respect to improvement of intestinal immunity in burned mice.

[Elucidation of structures and functions through Peyer's patches of responsible carbohydrate chains in intestinal immune system modulating polysaccharides from Japanese medicinal herbs].

Carbohydrate chains in glycoconjugates play important roles in various life phenomena, and there are numerous types of recognition system for carbohydrate chains due to carbohydrate-lectin interactions/carbohydrate-carbohydrate interactions in all higher life forms. It has been proposed that macromolecular polysaccharides isolated from plants, marine organisms, or fungi cross-interact with known and unknown recognition systems in mammals to express their pharmacological activities. Therefore the elucidation of carbohydrate structures related to the activities and functions of these polysaccharide molecules will lead us to utilize the related information in the development of novel carbohydrate-based drugs and functional foods for human health care. Peyer's patches present in the upper intestinal tract play important roles as inductive sites for both protective IgA production and immune tolerance induction in mucosal and systemic immune systems. Dysfunction of the immunocompetent cells of Peyer's patches is thought to induce allergic/autoimmune diseases and down-regulation of the protective system against infectious agents on mucosal sites. We have isolated several Peyer's patch cell-modulating polysaccharides from medicinal herbs used in traditional Japanese herbal remedies, and they have been assumed to comprise the responsible carbohydrate chains with oligosaccharide sizes for expression of modulating activity. Accumulation of knowledge on the structures and functions of these responsible carbohydrate chains in polysaccharide molecules is believed to be important for the development of methodology for logically factitious regulation of functions of immunocompetent cells in Peyer's patches. This review deals with recent results of our study on the structural clarification of responsible carbohydrate chains in modulating polysaccharides against functions of immunocompetent cells in Peyer's patches.

Effects of diets supplemented with organic acids and nucleotides on growth, immune responses and digestive tract development in weaned pigs.

Sixty-eight (Experiment 1, 46 days feeding) and sixteen (Experiment 2, 21 days feeding) 21-days-old weaned pigs were allotted to four dietary treatments including control, 0.6% organic acids (OA), 0.1% nucleotides (NA) and 0.6% OA plus 0.1% NA for determining the dietary effects. In Experiment 1, OA enhanced peripheral blood mononuclear cells proliferation on day 28 and 46. The plasma immunoglobulin (Ig) A level was elevated by OA (p < 0.06) and NA (p < 0.07), respectively. In Experiment 2, NA increased plasma IgM level, and had an interactive effect with OA on ileal Peyer's patches and mesenteric lymph node lymphocyte proliferation, bile and plasma IgA levels, and jejunal crypt depth. NA elevated gastric pepsin and jejunal alkaline phosphatase activities, however, decreased ileal aminopeptidase N, sucrase or maltase activity. These results suggest that OA and NA have synergistically enhanced the gut-associated lymphocyte responses and NA modulates the digestive tract development of weaned pigs.

Influence of the probiotic Bacillus cereus var. toyoi on the intestinal immunity of piglets.

In a feeding trial, sows and piglets were fed with the probiotic bacterium Bacillus cereus var. toyoi as a feed additive, and the effects on immune cell populations were examined. The development of the gut immune system was determined for piglets at the ages of 14, 28, 35 and 56 days post partum. Tissue samples of the Jejunum and the continuous Peyer's patch were used for enumeration of intraepithelial lymphocyte populations by fluorescence activated flow cytometry and fluorescence microscopy. Both independent methods of investigation led to similar results: the population of intraepithelial CD8+ T cells was significantly enhanced in the probiotic group piglets (p< or =0.05), and the numbers of gammadelta T cells tended to be higher in the intestinal epithelium (p<0.1) at the time of weaning (day 28). Lamina propria lymphocytes were also influenced by the treatment. Application of B. cereus var. toyoi resulted in significantly more CD25+ lymphocytes and gammadelta T cells in the probiotic group post-weaning. The occurrence of pathogenic Escherichia coli serogroups was also less frequent in the feces of piglets from the probiotic group. The finding that the CD8+ T cell population in the intestinal mucosa showed changes on day 28 indicated that the influence of B. cereus var. toyoi supplementation on the intestinal immune system started before weaning, an observation supported by changes in the intestinal microflora observed during the suckling-period. The results suggest that feeding of B. cereus var. toyoi to sows may result in beneficial effects on piglet health status independent of their feed supplementation.

Influence of adding fish oil to parenteral nutrition on gut-associated lymphoid tissue.

BACKGROUND: Lack of enteral nutrition reduces gut-associated lymphoid tissue (GALT) mass and function, a mechanism underlying the increased morbidity of infectious complications in severely injured or critically ill patients. Strategies to restore parenteral nutrition (PN)-induced changes of GALT mass and function have been pursued. However, the influences of adding fish oil to PN on gut immunity remain to be clarified. METHODS: Male Institute of Cancer Research (ICR) mice (n = 50) were randomized to 4 groups: ad libitum chow (chow), fat free PN (fat (-)-PN), PN + fish oil (FO-PN), and PN + safflower oil (SO-PN). The PN groups were given isocaloric and isonitrogenous PN solutions. The FO- and SO-PN groups received 20% of total calories from fat emulsions. After 5 days of feeding, lymphocytes from Peyer's patches (PPs), the intraepithelial space (IE), and the lamina propria (LP) of the entire small intestine were isolated. GALT lymphocyte numbers and phenotypes (CD4+, CD8+, alphabetaTCR+, gammadeltaTCR+, B220+ cells) were determined. Immunoglobulin A (IgA) levels of small intestinal washings were also measured by enzyme-linked immunosorbent assay. Another set of mice (n = 24) was used to determine plasma fatty acid compositions after feeding. RESULTS: Lymphocyte numbers from PPs and the LP and intestinal IgA levels were significantly lower in the PN groups than in the chow group, with no significant differences between any 2 PN groups. The FO- and SO-PN groups showed moderate recovery of IE cell numbers compared with the fat (-)-PN group. Omega-3 and omega-6 fatty acid levels were increased with fish and safflower oil additions, respectively, compared with the fat (-)-PN group. CONCLUSIONS: Adding fish oil to PN does not exacerbate PN-induced GALT changes but rather partially reverses these changes, with increased plasma omega-3 fatty acid levels.

Capsicum ethanol extracts and capsaicin enhance interleukin-2 and interferon-gamma production in cultured murine Peyer's patch cells ex vivo.

We investigated the effects of red pepper (Capsicum annuum Lin.) extracts (capsicum extract) and its main pungent capsaicin on T helper 1 (Th1) and 2 (Th2) cytokine production in cultured murine Peyer's patch (PP) cells in vitro and ex vivo. Direct administration of capsicum extract (1 and 10 mug/ml) and capsaicin (3 and 30 muM) resulted in suppression of interleukin (IL)-2, interferon (IFN)-gamma, IL-4 and IL-5 production. In an ex vivo experiment using PP cells removed from the mice after oral administration of capsicum extract (10 mg/kg/day for 4 consecutive days), IL-2, IFN-gamma and IL-5 increased in response to concanavalin A (Con A). Oral administration of 3 mg/kg/day capsaicin, one active constituent of the extract, also enhanced IL-2, INF-gamma and IL-4 production in response to Con A stimulation but did not influence the production of IL-5. Orally administered capsazepine (3 mg/kg/day), a selective transient receptor potential vanilloid 1 (TRPV1) antagonist, slightly enhanced IL-2 production also irrespective of Con A stimulation. The capsaicin-induced enhancement of both IL-2 and IFN-gamma production was not reduced by oral administration of capsazepine (3 mg/kg/day), suggesting a TRPV1 receptor-independent mechanism. Flow cytometric analysis revealed that the population of CD3(+) cells in the PP cells was significantly reduced while CD19(+) cells increased after oral administration of capsicum extract (1 and 10 mg/kg/day) and capsaicin (0.3 and 3 mg/kg/day). Capsazepine (3 mg/kg/day) weakly but significantly reversed these effects. Orally administered capsicum extract and capsaicin did not change the T cell subset (CD4(+) and CD8(+)), Th1 (IFN-gamma(+)) and T2 (IL-4(+)) ratio. These findings indicate that capsicum extract and capsaicin modulate T cell-immune responses, and their immunomodulatory effects on murine PP cells are partly due to both TRPV1-dependent and -independent pathway.

Toll-like receptor 2-dependent activation of monocytes by Spirulina polysaccharide and its immune enhancing action in mice.

We reported previously that a high molecular weight polysaccharide fraction (Immulina) from Spirulina was a potent activator of NF-kappa B and induced both IL-1 beta and TNF-alpha mRNAs in THP-1 human monocytes. In the present study, we show that NF-kappa B activation by Immulina is suppressed by antibodies to CD14 and TLR2 but not by antibodies to TLR4. Similarly, NF-kappa B directed luciferase expression was enhanced by Immulina treatment when cells were co-transfected with vectors expressing proteins supporting TLR2- (CD14 and TLR2) but not TLR4-(CD14, TLR4, and MD-2) dependent activation. Mice that consumed a chemically defined chow mixed with an extract containing Immulina exhibited changes in several immune parameters. The ex vivo production of IgA and IL-6 from Peyer's patch cells was enhanced 2-fold and interferon-gamma production from spleen cells was increased 4-fold in Immulina-treated mice. The enhanced production of these factors was most notable with mice that had consumed this extract for 4 or 5 days. These studies shed light on how Immulina activates cells of the innate immune system and suggests that oral consumption of this polysaccharide can enhance components within both the mucosal and systemic immune systems.

Effect of vitamin E supplementation on lymphocyte distribution in gut-associated lymphoid tissues obtained from weaned piglets.

Fifteen piglets were used to determine the effect of vitamin E supplementation on the number of CD4-immunoreactive (CD4+) T-lymphocytes, CD8-immunoreactive (CD8+) T-lymphocytes and IgA-immunoreactive (IgA+) B-lymphocytes per follicle in the Peyer's patch of distal ileum and the mesenteric lymph nodes of weaned piglets. Piglets, following a 3-day adaptation period after weaning, were assigned to one of three experimental groups: control (no vitamin E supplementation), vitamin E supplementation of 100 mg/kg of diet and vitamin E supplementation of 300 mg/kg of diet. Supplementation of vitamin E lasted for a period of 36 days. The basal diet contained 80 mg alpha-tocopherol/kg of diet. All piglets were killed at day 39 after weaning and samples of the distal ileum and adjacent mesenteric lymph nodes were collected. The number of cells for each lymphocyte subset was counted in the Peyer's patch and the mesenteric lymph nodes follicles, in cryostat sections processed for immunohistochemistry. Results showed that vitamin E supplementation (300 mg/kg diet) of piglets caused an increase (P < 0.05) in the number of IgA+ B-lymphocytes in the Peyer's patch, but not in the mesenteric lymph nodes, compared with the corresponding values in control animals. Vitamin E supplementation had no effect (P > 0.05) on the number of CD4+ and CD8+ T-lymphocytes in the follicles of the Peyer's patch and the adjacent mesenteric lymph nodes. Thus, vitamin E had relatively minor effects on distribution of the major immunocompetent cells in the gut. The numbers of CD4+ and CD8+ T-lymphocytes as well as IgA+ B-lymphocytes per follicle were higher by 26-77% (P < 0.05) in the mesenteric lymph nodes than the corresponding values in the Peyer's patch.

Glutamine supplementation influences immune development in the newly weaned piglet.

A study was conducted to determine changes that occur in immune function during the early post-weaning period and the effect of supplementing glutamine (gln, 4% w/w) to the weaning diet of piglets. Dutch-Landrace piglets (n=10/group) were killed prior to weaning (21 d) or randomized to one of two nutritionally complete weaning diets with or without gln. With age there was an increased ability of peripheral blood mononuclear cells (PBMC) and mesenteric lymph nodes (MLN) cells to proliferate (rate of (3)H-thymidine uptake) to T cell mitogens (P<0.05). PBMC from older piglets produced less of a Th-1 type response after stimulation (P<0.05). Adding gln to the weaning diet significantly (P<0.05) modified immune cells in the MLN, in a potentially beneficial manner (with respect to mucosal infections) by preventing an increase in antigen naïve CD4+ cells, increasing the proliferative response to pokeweed mitogen and supporting a Th-1 type cytokine response after T cell (phytohemagglutinin) stimulation.

Glutamine supplementation enhances mucosal immunity in rats with Gut-Derived sepsis.

OBJECTIVE: Supplemental glutamine (Gln) has been demonstrated to improve the immunologic response and reduce mortality in rodents with sepsis. However, the effects of Gln on gut-associated lymphoid tissue function after infection and sepsis are not clear. We investigated the effects of Gln-supplemented diets before sepsis, Gln-enriched total parenteral nutrition (TPN) after sepsis, or both on the intestinal immunity in rats with gut-derived sepsis. METHODS: Male Wistar rats were assigned to control and four experimental groups. The control and experimental groups 1 and 2 were fed a semi-purified diet; in experimental groups 3 and 4, part of the casein in the diets was replaced with Gln. After feeding rats the respective diets for 10 d, sepsis was induced by cecal ligation and puncture (CLP) in the experimental groups, whereas the control group underwent a sham operation; at the same time, the internal jugular vein of all rats was cannulated. All rats were maintained on TPN for 3 d. The control group and groups 1 and 3 were infused with conventional TPN, and groups 2 and 4 were given a TPN solution supplemented with Gln, which provided 25% of total amino acid nitrogen. All rats were killed 3 d after the sham operation or CLP. Intestinal immunoglobin A levels, total lymphocyte yields, and lymphocyte subpopulations in Peyer's patches were analyzed. RESULTS: Total Peyer's patch lymphocyte numbers were significantly higher in the Gln-supplemented groups than in the control group. Distributions of CD3+ and CD4+ in group 1 were significantly lower than those in the control group, whereas no differences were observed among the control and Gln-supplemented groups. Plasma immunoglobulin A levels were higher in the Gln-supplemented groups than the control group and group 1. Intestinal immunoglobulin A levels were significantly higher in groups 2 and 4 than in the control group and group 1. CONCLUSIONS: Preventive use of a Gln-supplemented enteral diet before CLP or intravenous Gln supplementation after CLP have similar effects in promoting proliferation of total lymphocyte in gut-associated lymphoid tissue, enhancing IgA secretion, and maintaining T-lymphocyte populations in Peyer's patches. Gln administered before and after CLP did not seem to have a synergistic effect on enhancing mucosal immunity in rats with gut-derived sepsis.