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Low colostral IgA associated with cow's milk allergy.

During a nutritional study of 198 infants, seven became allergic to cow's milk. The seven infants showed acute cutaneous manifestations during cow's milk challenge tests in hospital and six had increased levels of IgE cow's milk-specific antibodies. Neither in the development of the levels of immunoglobulins G, A and M, nor in that of the cow's milk-specific antibodies of these isotypes did these seven patients differ from the remaining infants. Beta-lactoglobulin content and levels of cow's milk-, and beta-lactoglobulin-specific antibodies and of immunoglobulins A, G and M were measured in samples of colostrum and milk from the mothers of the seven infants with cow's milk allergy and from a comparison group (non-atopic mothers of non-atopic infants). The milk of the mothers whose infants became allergic to cow's milk contained less IgA through the lactation: 95% confidence intervals of the groups did not overlap. The difference was most marked in the colostrum. All other measurements were similar in the two groups. This suggests that an infant is more likely to develop cow's milk allergy if the mother's colostrum had a low total IgA content.

Animals↗

Breast feeding modifies production of SIgA cow's milk-antibodies in infants.

One hundred and fifty-eight healthy mature newborns were divided into 3 groups according to their risk of allergy: Group A, no risk (n = 62), group B, low risk (n = 30) and Group C, high risk (n = 66). Saliva was collected at birth and after 3 and 6 months. SIgA anti-casein and anti-beta-lactoglobulin were determined by a direct ELISA technique. The highest concentrations of secretory antibodies were measured after birth. After 3 months, breast fed infants had lower salivary SIgA anti-casein concentrations than the group receiving cow's milk (p less than 0.01). The effect of breast-feeding was seen even after a nursing period of only 3 weeks. Infants without risk of allergy fed cow's milk exclusively had higher SIgA anti-casein (p less than 0.03) and anti-beta-lactoglobulin concentrations than low risk infants at the age of 6 months. These data show a modifying effect of breast feeding on salivary SIgA production against cow's milk protein.

Animals↗

A comparison of secretory antibodies in breast-fed and formula-fed infants over the first six months of life.

In the present study salivary IgA, anti-Escherichia coli, anti-beta-lactoglobulin and anti-poliovirus type 1 IgA and IgM in serum and saliva were evaluated longitudinally in 13 breast-fed and 14 formula-fed infants over the first six months of life. Salivary IgA was quantified by electroimmunodiffusion; specific IgA and IgM antibodies were determined in serum and saliva by ELISA. Salivary IgA was significantly lower at age one month in breast-fed compared with formula-fed infants but in breast-fed infants salivary IgA increased with age and was significantly higher at six months than at one month. In both groups of infants, at the age of six months, salivary IgA levels were significantly lower than in adult controls. No significant differences in secretory anti-E. coli were observed between the two groups of infants. Salivary anti-poliovirus IgA and IgM antibodies increased transiently only to disappear in most babies at age six months, while anti-beta lactoglobulin IgA and IgM, present in saliva at all ages, showed a wide scatter. No important differences in specific serum IgA or IgM antibodies were observed either between the groups or at different times within the groups.

Age Factors↗

IgA bovine serum albumin antibodies are increased in newly diagnosed patients with insulin-dependent diabetes mellitus, but the increase is not an independent risk factor for diabetes.

We studied the significance of antibodies to bovine serum albumin (BSA) as a risk factor for insulin-dependent diabetes mellitus (IDDM) in a case-control setting. IgA and IgG antibodies to BSA and ovalbumin were measured from sera of 104 patients with newly diagnosed IDDM and of 111 matched controls by enzyme-linked immunosorbent assay. Patients with diabetes had significantly higher levels of IgA antibodies to BSA (p = 0.003); IgG antibodies also tended to be higher (p = 0.08). Levels of IgA antibodies to ovalbumin were similar in the patients and controls, but IgG antibodies were higher in controls (p = 0.02). When antibodies to BSA, beta-lactoglobulin, whole cow's milk and islet cell antibodies were studied as risk determinants of IDDM in a multivariate, logistic regression analysis, IgA antibodies to beta-lactoglobulin and to cow's milk were independently associated with the risk (p = 0.037 and 0.048, respectively), while antibodies to BSA were not a significant risk factor. The results question the role of BSA as a cross-reacting antigen with pancreatic beta-cell surface proteins in the aetiology of IDDM.

Animals↗

Dietary factors influencing levels of food antibodies and antigens in breast milk.

The effect of a milk-free diet during late pregnancy and lactation, on levels of cows' milk specific antibodies and antigens in breast milk, was examined. Women with an allergic condition (defined as atopic) were randomly allocated to a milk-free diet (n = 10) or an unrestricted diet (n = 12). Twelve non-atopic women followed an unrestricted diet. A significant reduction (p < 0.001) in beta-lactoglobulin-specific immunoglobulin A and alpha-casein-specific immunoglobulin A levels was observed over the first 5 days in all groups. The mean level of beta-lactoglobulin antigens in breast milk from women who adhered strictly to the milk-free diet was significantly lower than the levels of the atopic group on the unrestricted diet (p < 0.02). The allergy incidence in the infants born in the atopic diet group was significantly lower compared with that of the atopic group on the unrestricted diet.

Analysis of Variance↗

Difference between bacterial and food antigens in mucosal immunogenicity.

The mucosa-associated lymphoid tissue may deviate from its systemic counterpart in being able to discriminate between microbial and nonmicrobial antigens. To study this, the systemic and mucosal antibody responses to bacterial and food antigens were followed in parallel in female rats during two pregnancies and lactation periods. Germfree rats were monocolonized with an Escherichia coli O6K13H1 strain, and their diet was switched to pellets containing large amounts of ovalbumin and beta-lactoglobulin. Antibodies against O6 lipopolysaccharide already appeared in serum and bile 1 week after colonization, and those against type 1 fimbriae appeared a few weeks later. Serum immunoglobulin G antibodies against the E. coli enzyme beta-galactosidase were found in moderate titers in all rats after 16 weeks of exposure. In contrast, few rats had detectable antibody levels against the dietary proteins ovalbumin and beta-lactoglobulin in serum or bile even after 16 weeks of exposure. In the milk, antibodies against E. coli beta-galactosidase and type 1 fimbriae reached the highest titers, while moderate titers were found against the food antigens and against O6 lipopolysaccharide. The difference in immune reactivity against bacterial versus dietary antigens was not likely due to insufficient amounts of the latter reaching lymphoid tissue, since (i) uptake studies indicated that ovalbumin was more efficiently taken up than endotoxin and (ii) the same difference in antigenicity between ovalbumin and E. coli was seen after immunization directly into Peyer's patches. We therefore suggest that a prerequisite for a strong mucosal antibody response is that the antigen be encountered by the gut-associated lymphoid tissue within microorganisms capable of stimulating antigen presentation.

Animals↗

Early induction of secretory immunity in infancy: specific antibody in neonatal breast milk.

Neonatal breast milk from 50 babies aged between 2 and 39 days was studied for the presence of antibody to the cows' milk protein beta lactoglobulin. Specific IgA antibody and specific secretory antibody to beta lactoglobulin were detectable towards the end of the second week of life in milk secreted by neonates fed cows' milk formula. Specific antibody concentrations were independent of total IgA concentrations. Babies receiving little or no cows' milk protein did not produce antibody in neonatal breast milk. Antigen specific mucosal immune responses develop in tissues distant from the site of primary mucosal exposure by the end of the second week of life in term human neonates, suggesting that prophylactic immunisation against enteric or other mucosal pathogens within a few days of birth may provide antibody responses in secretions, which may protect against mucosal infection.

Aging↗

Effect of gliadin and other food peptides on expression of MHC class II molecules by HT-29 cells.

Expression of major histocompatibility (MHC) class II molecules by enterocytes is known to be enhanced in coeliac disease and other disorders characterised by intestinal inflammation--an effect thought to be mediated via intestinal lymphocytes. To investigate if food peptides can exert direct effects on class II expression, the influence of gliadins, casein, and beta lactoglobulin on an intestinal epithelial cell line (HT-29) was examined in the absence of immune cells. Class II expression was determined by flow cytometry and immunofluorescence microscopy using antibodies against the beta chain of all products of the gene subregions DR, DQ, and DP. MHC expression was low in HT-29 cells but could be stimulated by interferon gamma. Tryptin digested gliadin had no effect on class II expression. In the presence of interferon gamma, however, it was able to amplify MHC class II expression to mean (SEM) 150 (4)%. Casein exerted a similar effect (160 (14)%), but undigested gliadin, tryptin digested casein, and beta lactoglobulin had no influence. The observations suggest that within the concert of cytokine mediated interactions between enterocytes and lymphocytes, some dietary peptides could upregulate the presentation of food antigens, leading to a more efficient stimulation of lymphocytes, which in the case of coeliac disease might result in damage to the enterocytes.

Adenocarcinoma↗

The molecular manipulation of milk composition.

The introduction of cloned genes into the mouse germ line is now routine. Although more difficult technically, gene transfer has been accomplished in farm animals and offers the potential for genetic improvement. In this regard, we have been investigating the use of transgenic animals as production vehicles for high value proteins in milk. We have shown that DNA sequences derived from the gene encoding sheep beta-lactoglobulin mediate efficient and specific expression in the mammary gland. A fusion gene comprising beta-lactoglobulin sequences and those encoding antihemophilic human factor IX has been constructed. This construct has been introduced into sheep; it is expressed in the mammary gland, and the corresponding protein is secreted into milk.

Animal Husbandry↗

Viscoelasticity of a protein monolayer from anisotropic surface pressure measurements.

We present a method to completely characterize the viscoelasticity of Langmuir monolayers. In contrast to existing techniques, both the compression and shear moduli are determined at the same time, in a single experiment and with a standard apparatus. This approach relies on the measurement of anisotropy in the surface pressure: the tension is measured in orientations perpendicular and parallel to the compression direction. We apply this technique to the study of beta-lactoglobulin spread monolayers, a system that has been shown to develop a shear modulus at high concentration. Beta-lactoglobulin monolayers are interesting both because of their importance in food science and because they exhibit universally slow dynamical behavior that is not fully understood. Our results confirm that the compressional modulus dominates the total viscoelastic response and also provide a complex shear modulus, emerging above a critical concentration. We are able to describe how each of the dynamical response moduli is related to the surface concentration and to the equilibrium osmotic pressure.

Algorithms↗

Neuroimmune interactions in guinea pig stomach and small intestine.

Enteric neuroimmune interactions in gastrointestinal hypersensitivity responses involve antigen detection by mast cells, mast cell degranulation, release of chemical mediators, and modulatory actions of the mediators on the enteric nervous system (ENS). Electrophysiological methods were used to investigate electrical and synaptic behavior of neurons in the stomach and small intestine during exposure to beta-lactoglobulin in guinea pigs sensitized to cow's milk. Application of beta-lactoglobulin to sensitized preparations depolarized the membrane potential and increased neuronal excitability in small intestinal neurons but not in gastric neurons. Effects on membrane potential and excitability in the small intestine were suppressed by the mast cell stabilizing drug ketotifen, the histamine H(2) receptor antagonist cimetidine, the cyclooxygenase inhibitor piroxicam, and the 5-lipoxygenase inhibitor caffeic acid. Unlike small intestinal ganglion cells, gastric myenteric neurons did not respond to histamine applied exogenously. Antigenic exposure suppressed noradrenergic inhibitory neurotransmission in the small intestinal submucosal plexus. The histamine H(3) receptor antagonist thioperamide and piroxicam, but not caffeic acid, prevented the allergic suppression of noradrenergic inhibitory neurotransmission. Antigenic stimulation of neuronal excitability and suppression of synaptic transmission occurred only in milk-sensitized animals. Results suggest that signaling between mast cells and the ENS underlies intestinal, but not gastric, anaphylactic responses associated with food allergies. Histamine, prostaglandins, and leukotrienes are paracrine signals in the communication pathway from mast cells to the small intestinal ENS.

Anaphylaxis↗

Permeability of intestinal capillaries to small molecules.

The permeability of capillaries in the isolated, vascularly perfused cat ileum was studied with the double-indicator diffusion technique. The molecules studied were raffinose, inulin, and beta-lactoglobulin A. Plasma flow through the tissue was increased by local intra-arterial infusions of isoproterenol (a selective mucosal vasodilator) or adenosine (a dilator of the muscularis vasculature). The permeability-surface area product (PS) for all solutes increased with increasing plasma flow. At the highest plasma flow acquired with isoproterenol, PS values for raffinose, inulin, and beta-lactoglobulin A were approximately 40, 20, and 4 ml.min-1.100 g-1, respectively. These data suggest that capillaries in the mucosa-submucosa of the ileum are at least five to seven times more permeable to raffinose and inulin than has been reported for skeletal muscle capillaries. Selective vasodilation of the muscularis region with adenosine reduced PS values for raffinose and inulin at each flow rate studied. The data are consistent with equivalent pore radii of approximately 60 A for capillaries in the mucosa-submucosa and 40 A in the muscularis of the small intestine.

Animals↗

Neuroimmune communication in the submucous plexus of guinea pig colon after sensitization to milk antigen.

We investigated electrical and synaptic behavior of neurons in the colonic submucous plexus during exposure to beta-lactoglobulin in guinea pigs sensitized by ingestion of milk. Microelectrodes were used to record electrical and synaptic behavior in neurons from milk-sensitized and nonsensitized age-matched animals during exposure to beta-lactoglobulin. Neurons in sensitized animals were hyperexcitable. Application of the histamine H2 antagonist cimetidine reversed the hyperexcitability, suggesting endogenously released histamine as one of the responsible factors. Antigenic exposure suppressed stimulus-evoked nicotinic cholinergic fast excitatory postsynaptic potentials (EPSPs). This was blocked by the selective histamine H3 antagonist burimamide. Suppression of the EPSPs resulted from presynaptic inhibition of acetylcholine release. Increased neuronal excitability and suppression of synaptic transmission was only found in milk-sensitized intestine, not in the intestine from age-matched nonsensitized animals. We concluded that signaling from mucosal mast cells to the enteric nervous system is important in colonic defense against antigenic threats. Histamine is a paracrine messenger in the communication network.

Acetylcholine↗

Paracellular transport of avidin saturated or not with biotinylated cobalamin through Caco-2 cell epithelium monolayer.

BACKGROUND: The cationic charge of molecules may promote their uptake across epithelia, which are rich in brush border anionic sites. The transport of unsaturated avidin and avidin saturated with a biotinylated compound was investigated across Caco-2 adenocarcinoma cell with fetal enterocyte phenotype. METHODS: The unsaturated avidin and avidin saturated with either biotin or a biotinyl-cobalamin conjugate (biotinyl-Cbl) were iodinated to follow their transport through the cell monolayer. Their apparent permeability coefficient (Papp) and transepithelial pathway were determined and compared to those for control radiolabeled markers [3H]-mannitol, [125I]-beta-lactoglobulin and [57Co]-cobalamin/intrinsic factor (Cbl/IF). RESULTS: The Papp of [125I]-avidin estimated at 2.8 x 10(-7) +/- 0.08 cm/s was close to that for mannitol that uses paracellular pathway. The binding of biotin or biotin conjugate to avidin enhanced its tetrameric conformation. The Papp for [125I]-avidin/biotin and [125I]- avidin/biotinyl-Cbl were respectively increased by 2-fold, compared to that for [125I]-avidin and 4-fold, compared to that for [125I]-beta-lactoglobulin and [54Co]-Cbl/IF. The protein was not accumulated in the cell and was found in intact form in the basolateral side, after its transport across the monolayer. Chloroquine (0.66 micromol/ml) did not significantly decrease the Papp for [125I]-avidin/biotinyl-Cbl. Conversely it decreased by 80% the Papp for Cbl/IF, that uses transepithelial pathway. CONCLUSIONS: Avidin (either saturated or not with biotin and biotinyl-Cbl) was able to cross the monolayer of Caco-2 cell line through a paracellular pathway. This study pointed out the interest for using this protein as a shuttle for increasing the transport rate of biotinylated compounds through fetal epithelial barriers.

Avidin↗

An increase in circulating IgA antibodies to gliadin in IgA mesangial glomerulonephritis.

In IgA glomerulonephritis (GN), the pathogenic role of IgA is well documented, but the specificity of these IgA is unknown. Cases of celiac disease associated with IgA GN have been reported and led us to investigate the role of gliadin sensitivity. We measured IgA, IgG and IgM antibodies to gliadin, beta-lactoglobulin and ovalbumin by ELISA (results expressed as optical density; OD) in 27 patients with primary IgA GN, 14 with membranous GN (MGN), 21 with idiopathic nephrotic syndrome (INS) and 21 healthy controls. The normal value for antigliadin IgA was less than 0.650 OD. 19/27 patients with IgA GN had a raised level versus 2/14 in MGN and 2/21 INS (p less than 0.001: IgA GN vs. MGN, INS and controls). Antibodies to beta-lactoglobulin were rarely found and were not more frequent in IgA GN. Cross-reactivity with reticulin was investigated in 16 patients who were serum-positive for IgA antigliadin: no reticulin antibodies were detected by immunofluorescence. Antigliadin IgA are of diagnostic value for distinguishing IgA GN from other GN, with a sensitivity of 70%, a specificity of 89%, a positive predictive value of 83% and a negative one of 79%.

Adult↗

Tolerance to orally induced anaphylactic sensitisation to cow's milk proteins and patency of the intestinal mucosa in the neonatal guinea pig.

Guinea pigs fed cow's milk within 2 days of birth may acquire tolerance to anaphylactic sensitisation to cow's milk proteins. Appreciable amounts of beta-lactoglobulin may be detected in their blood, in contrast with animals receiving cow's milk later than this age. Uptake of beta-lactoglobulin from the gut to circulation ceases after the 6th postnatal day. These findings could be relevant to cot death in the human infant should the anaphylactic hypothesis be substantiated.

Anaphylaxis↗

The effects of exclusion of dietary egg and milk in the management of asthmatic children: a pilot study.

Current understanding of the use of exclusion diets in the management of asthma in children is limited and controversial. The aim of this study was to examine the effects of excluding eggs and milk on the occurrence of symptoms in children with asthma and involved 22 children aged between three and 14 years clinically diagnosed as having mild to moderate disease. The investigation was single blind and prospective, and parents were given the option of volunteering to join the 'experiment' group, avoiding eggs, milk and their products for eight weeks, or the 'control' group, who consumed their customary food. Thirteen children were recruited to the experimental group and nine to the control group. A trained paediatrician at the beginning and end of the study period assessed the children. A seven-day assessment of food intake was made before, during and immediately after the period of dietary intervention in both groups. A blood sample was taken from each child for determination of food specific antibodies and in those children who could do so, the peak expiratory flow rate (PEFR) was measured. Based on the recommended nutrient intake (RNI), the mean percentage energy intake of the children in the experimental group was significantly lower (p < 0.05) in the experimental group. After the eight-week study period and compared with baseline values, the mean serum anti-ovalbumin IgG and anti-beta lactoglobulin IgG concentrations were statistically significantly reduced (p < 0.05) for both in the experimental group. In contrast, the values for anti-ovalbumin IgG in the control group were significantly increased and those for anti-beta lactoglobulin IgG were practically unchanged. The total IgE values were unchanged in both groups. Over the study period, the PEFR in those children in the experimental group able to perform the test was significantly increased, but no such change was noted in the children in the control group who could do the test. These results suggest that even over the short time period of eight weeks, an egg- and milk-free diet can reduce atopic symptoms and improve lung function in asthmatic children.

Adolescent↗

IgD in human colostrum.

Simultaneous colostrum (C) and plasma samples (P) from 14 women, 1 to 5 days postpartum, were examined. Total IgD and specific IgD antibodies to beta-lactoglobulin, bovine serum albumin, Bermuda grass, and alpha-gliadin were measured by solid phase radioimmunoassay. The geometric mean concentrations of IgD were 35.8 (range 2.2-410) micrograms/dl for colostrum and 591.3 (range 72-4100) micrograms/dl for plasma. Six subjects had a specific IgD antibody C/P ratio more than 10-fold greater than the total IgD C/P ratio, suggesting enhancement of antibody to a specific antigen in the mammary gland. All six had C/P ratios suggestive of local enhancement of IgD antibody to Bermuda grass, and two met this criterion for enhancement of IgD antibodies to beta-lactoglobulin, bovine serum albumin, or alpha-gliadin. Specimens for these studies were obtained during the peak grass pollen season. Seventeen additional subjects were studied to compare total IgD in colostrum and plasma with total IgG and serum albumin. The mean C/P ratio for IgD (0.055 +/- 0.015) exceeded the C/P ratio for total IgG (0.015 +/- 0.003) or total albumin (0.020 +/- 0.002). For 14 of 17 subjects the colostrum/plasma ratio for IgD exceeded the C/P ratio for albumin or IgG. Data were transformed logarithmically and correlation coefficients calculated. For albumin versus IgG in colostrum, there was a strong correlation, r = 0.865, p = 0.001. This was different from albumin versus IgD, r = 0.489, p = 0.046 and from IgD versus IgG, r = 0.556, p = 0.020. These analyses support a different mechanism of entry of IgD into milk compared to IgG or albumin.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibody Formation↗