Inhibition of verotoxin (VT) 2 absorption into systemic blood from intestine by repeated administration of bovine immune colostral antibody against VT2 in mice.

BACKGROUND/PURPOSE: Whether absorption of verotoxin (VT) 2 from the intestine in mice is inhibited by administration bovine immune colostral antibody against VT2 was investigated. METHODS: Three-week-old mice were administered VT2 solution at 477.8 ng/mL or 955.6 ng/mL, and bovine immune colostral antibody against VT2 was then administered three times. Whey without antibody against VT2 was administered to control mice. Serum levels of VT2 were measured by fluorescence enzyme immunoassay. RESULTS: Serum levels of VT2 in mice administered VT2 solution at 477.8 ng/mL and bovine immune colostral antibody against VT2 scarcely changed. By contrast, serum levels of VT2 in control mice increased and peaked 12 hours after administration. Peak values were 15.4 ± 5.04 ng/mL. Furthermore, serum levels of VT2 at 12 hours and 16 hours in control mice were significantly higher than in mice administered bovine colostral antibody against VT2. Serum levels of VT2 in mice administered antibody at 955.6 ng/mL showed no significant differences between repeated administration of bovine immune colostral antibody and controls. CONCLUSION: These results suggest that absorption of VT2 from the intestine was inhibited by repeated administration of bovine immune colostral antibody against VT2 at early stages of Escherichia coli O157:H7 infection, whereas VT2 in the intestine remained at low levels.

Interleukin-22 modulates gut epithelial and immune barrier functions following acute alcohol exposure and burn injury.

Interleukin-22 (IL-22) maintains gut epithelial integrity and expression of antimicrobial peptides Reg3ß and Reg3γ. Our laboratory has shown that acute alcohol/ethanol (EtOH) exposure before burn injury results in increased gut permeability, intestinal T-cell suppression, and enhanced bacterial translocation. Herein, we determined the effect of combined EtOH intoxication and burn injury on intestinal levels of IL-22 as well as Reg3ß and Reg3γ expression. We further examined whether in vivo restitution of IL-22 restores gut permeability, Reg3ß and Reg3γ levels, and bacterial load (e.g., gut bacterial growth) within the intestine after EtOH and burn injury. Male mice, ∼25g, were gavaged with EtOH (2.9 mg/kg) before receiving a ∼12.5% total-body-surface-area, full-thickness burn. Mice were immediately treated with saline control or IL-22 (1 mg/kg) by i.p. injection. One day after injury, there was a significant decrease in intestinal IL-22, Reg3ß, and Reg3γ expression along with an increase in intestinal permeability and gut bacterial load after EtOH combined with burn injury, as compared with sham injury. Treatment with IL-22 normalized Reg3ß and Reg3γ expression and attenuated the increase in intestinal permeability after EtOH and burn injury. Qualitatively, IL-22 treatment reduced the bacterial load in nearly half of mice receiving EtOH combined with burn injury. Our data indicate that IL-22 maintains gut epithelial and immune barrier integrity after EtOH and burn injury; thus, the IL-22/antimicrobial peptide pathway may provide a therapeutic target for the treatment of patients who sustain burn injury under the influence of EtOH.

Short communication: Addition of varying amounts of sodium bicarbonate to colostrum replacer: effects on immunoglobulin G absorption and serum bicarbonate in neonatal calves.

Fifty-two dairy calves were blocked by birth date and, within each block, randomly assigned to 1 of 4 treatments to investigate the effects of incremental levels of sodium bicarbonate (NaHCO(3)) on IgG metabolism. Treatments were (1) colostrum replacer (CR)+0 g of NaHCO(3) (control); (2) CR+15 g of NaHCO(3); (3) CR+30 g of NaHCO(3); or (4) CR+45 g of NaHCO(3). Calves were fed colostrum replacer (>200 g of IgG) in one feeding within 45 min of birth (0 h) and 2 L of milk replacer at 12, 24, 36, and 48 h. Only calves born in calving pens from multiparous cows with no dystocia were used in this study. Blood samples were taken at 0, 6, 12, 24, and 48 h postpartum, and serum was analyzed for IgG using radial immunoassay and bicarbonate using spectrophotometry. Feeding increasing levels of sodium bicarbonate had negative linear effects on IgG concentration, IgG apparent efficiency of absorption, and IgG area under the curve, primarily due to the effect of the highest dose of NaHCO(3) (45 g). Sodium bicarbonate treatments had no effect on serum bicarbonate concentration. However, area under the curve of serum bicarbonate increased linearly with the amount of NaHCO(3) fed.

Immunomodulation. Part IV: Glutamine.

Glutamine, a nonessential amino acid that appears to be conditionally essential during periods of physiologic stress, plays important physiologic roles in the immune system. However, neither enteral nor parenteral glutamine supplementation makes a difference in the rate of systemic infection or of NEC in very low birth weight infants. Thus, the search for agents to enhance the neonate's immune system and to serve as safe and effective adjuvants to antibiotics continues. Part V, the final article in this immunomodulation series, will explore the use of probiotics to support the neonatal immune system.

Specific immunoglobulins in serum of newborn lambs fed with a single dose of colostrum containing anti-peroxidase IgG.

The apparent efficiency of absorption and the decrease of specific colostral IgG after its passage into the blood stream were determined in newborn lambs fed with a single dose of colostrum containing anti-peroxidase IgG at 30 minutes, 12 hours and 24 hours after birth. When colostrum was given at 30 minutes after birth, a value of 16.9+/-4.0 per cent of anti-peroxidase IgG ingested appeared in lamb circulation. This percentage was reduced to 9.8+/-0.8 per cent when the feeding was done at 12 hours after birth and no specific IgG was detected in lambs fed at 24 hours after birth. The concentration of anti-peroxidase IgG in lambs' serum declined quickly within 96 hours of age to about 48 per cent of the initial value, and afterwards the level decreased slowly reaching a value of 10 per cent at 32 days of age. This behaviour probably reflects the protein distribution and use of absorbed antiperoxidase IgG.

Gamma delta intraepithelial lymphocytes drive tumor necrosis factor-alpha responsiveness to intestinal iron challenge: relevance to hemochromatosis.

The dependence of intestinal epithelial cell (IEC) growth and differentiation on intraepithelial lymphocytes (IELs) expressing the gamma/delta (gamma delta) T-cell receptor (TCR), suggested a potential role for gamma delta + IELs in the regulation of iron absorption. We therefore examined the levels of hepatic iron and the IEL cytokine responses in C57BL/6J control and class I and TCR knockout lines (placed on a C57BL/6J genetic background) following the administration of supplemental dietary iron. The highest level of liver iron was found in the beta 2-microglobulin knockout (beta 2m-/-) mice followed by the TCR-delta knockout (TCR delta-/-) animals. TCR-alpha knockout (TCR alpha-/-) and control animals did not differ in their iron levels. Liver iron loading correlated inversely with the ability of the mice to generate an IEL tumor necrosis factor (TNF)-alpha response. These observations suggest a model in which IEC iron loading is communicated to IELs via the HFE class I protein. The result of this communication is the initiation of TNF-alpha release by gamma delta + IELs (sustained by macrophages and dendritic cells) contributing to the upregulation of ferritin expression and possibly to the normal maintenance of the IEC apoptotic pathway.

An immunoelectron microscopic investigation of colostral IgG absorption across the intestine of newborn calves.

The protein A-gold was used to examine the transport of colostral IgG from the lumen of the gut to the circulation in four newborn calves and one 24-hour-old calf. The absorptive enterocytes of the duodenum, jejunum and ileum were investigated five to 60 minutes after administering colostrum, and 24 hours after birth. In the newborn calves, an intracellular micropinocytotic transport of IgG molecules was dominant throughout the entire small intestine. The amount transported increased from the duodenum to the ileum. In addition, evidence of a selective, receptor-mediated transport of IgG during the first few hours of life was provided by the presence of bovine clathrin at the microvillous membrane of the duodenal and jejunal enterocytes, indicating the existence of specialised vesicles for transport, the so-called 'coated' vesicles. No sign of paracellular transport was detected. Intestinal closure was interpreted as a multifactorial event comprising the replacement of the fetal intestinal epithelial cells by more mature populations, the initial cessation of transport at the basal and lateral cell membrane of the absorptive enterocytes, and an increase in intracellular proteolytic activity by lysosomes.

Effect of bovine serum albumin on passive transfer of immunoglobulin G1 to newborn calves.

The molecular mechanism in the intestine of newborn calves that results in transfer of intact colostral immunoglobulin from the lumen to the circulation also is capable of transferring a variety of non-immunoglobulin macromolecules. If the capacity of this mechanism is limited, transfer of a large amount of non-immunoglobulin protein may interfere with transfer of immunoglobulin. In this experiment, efficiency of IgG1 transfer in newborn calves was reduced from 59 to 36% by the addition of bovine serum albumin (37 mg ml-1) to colostral whey, while the addition of a similar mass of amino acids in the form of acid hydrolyzed casein (37 mg ml-1) did not detectably alter IgG1 transfer. Reduced IgG1 absorption efficiency in calves fed colostrum with added bovine serum albumin is consistent with a limited capacity for the macromolecular transport mechanism in the intestine of newborn calves.

Intestinal absorption of immunologically intact macromolecules in germfree colostrum-deprived piglets maintained on total parenteral nutrition.

We have compared the neonatal absorption of anti-bovine gamma-globulin (BGG) antibody supplied in colostrum or saline in three groups of piglets born and maintained under different environmental conditions to determine the effect of these conditions on the cessation of intestinal absorption of macromolecules (anti-BGG antibody), termed "closure." An enzyme-linked immunosorbent assay was used to estimate the concentration of anti-BGG antibody in sera from each group of piglets. Three stages of macromolecular absorption through the piglet's intestine could be detected. The first stage is a nonselective massive absorption of macromolecules (in milligram levels) that lasts up to 3 days in germfree (GF) colostrum-deprived or conventional colostrum-fed piglets but up to 5 days in GF piglets maintained on total parenteral nutrition. In this stage, absorption was significantly (r = .05) higher in piglets fed anti-BGG serum with colostrum than in piglets fed anti-BGG serum without colostrum on GF day 0 (31.28% vs 15.59%) and GF-total parenteral nutrition day 3 (3.08% vs 0.11%). Thus, whenever there was the ability to absorb a massive amount of macromolecules, the sow colostrum had an enhancing affect. Although there was a minor effect of environmental or orally received stimuli in delaying closure, absorption of macromolecules decreased in all piglets maintained either parenterally or enterally after day 3. Thus, intestinal closure to massive absorption of macromolecules in piglets is primarily time (age)-dependent. The second stage is a selective absorption of immunoglobulins in much smaller quantities (microgram levels), inasmuch as absorption of 0.02% to 0.1% was determined in all 5-day-old piglets.(ABSTRACT TRUNCATED AT 250 WORDS)