Detection of specific IgE to human milk proteins in sera of atopic infants

Specific IgE (sIgE) for cow's milk proteins (CMP) have been reported to be present in blood sera of exclusively breast-fed infants. The aim of this study was to find whether the presence of sIgE to human milk proteins in the sera of exclusively breast-fed infants could explain the apparent detection of sIgE to CMP in infants that were never previously in contact with cow's milk. sIgE for human milk whey proteins were found in the blood sera of atopic infants, and these sIgE strongly cross-reacted with the corresponding CMP. In none of the sera examined were sIgE to bovine beta-lactoglobulin detected.     

Intestinal barrier function and cow's milk sensitization in guinea pigs fed milk or fermented milk

BACKGROUND: The respective effect of milk and fermented milks on intestinal barrier capacity and on sensitization to beta-lactoglobulin was studied using a guinea pig model of cow's milk allergy. METHODS: Guinea pigs were fed a control diet or the same diet supplemented with milk, fermented milk (Streptococcus thermophilus and Bifidobacterium breve), or dehydrated fermented milk. Intestinal barrier capacity to macromolecules was assessed in an Ussing chamber, and sensitization to cow's milk proteins was measured by systemic anti-beta-lactoglobulin immunoglobulin G1 titers and by intestinal anaphylaxis, the latter assessed by the beta-lactoglobulin-induced increase in short-circuit current of jejunal fragments (deltaIsc(beta-LG)). RESULTS: The electrical resistance of jejunum was similar in the four groups (approximately 80 omega/cm2) suggesting the same paracellular permeability. The transport of 14C-beta-lactoglobulin from mucosa to serosa was significantly decreased in the animals fed dehydrated fermented milk (403+/-131 ng / hr x cm2) compared with that in control animals or animals fed milk (767+/-250 ng / hr x cm2 and 749+/-475 ng / hr x cm2, respectively; p < 0.05). Milk fermentation did not modify native beta-lactoglobulin concentration but anti-beta-lactoglobulin immunoglobulin G1 titers were higher in fermented milk and dehydrated fermented milk (log10 titer = 2.86 and 2.79, respectively) than in guinea pigs fed milk (log10 titer = 2.5; p < 0.007). However, beta-lactoglobulin-induced intestinal anaphylaxis remained the same in the three groups (deltaIsc(beta-LG), 9.6+/-4.1 microA/cm2, 8.5+/-4.3 microA/cm2, and 8.5+/-3.4 microA/cm2 in milk-fed, fermented milk-fed, and dehydrated fermented milk-fed guinea pigs, respectively). CONCLUSIONS: The intestinal barrier capacity to milk proteins seems to be reinforced by dehydrated fermented milk, but milk and fermented milks are equally efficient in inducing cow's milk allergy in guinea pigs.     

Multiple food allergy: a possible diagnosis in breastfed infants

Six infants suspected of food allergy during breastfeeding were evaluated using prick tests, total IgE, RASTs and intestinal permeability measurements during fast and provocation with mother's milk. An elimination diet was undertaken in mothers, removing first cow's milk protein (CMP), then, when inefficient, all foods suspected on the clinical history or a positive prick test in the child, followed by oral challenges in mother's diet with the corresponding food. The sole CMP-free diet in mothers always proved insufficient. In four, an additional diet excluding two to three other foods cleared the symptoms. Oral provocations in mother's diet with those foods were positive in all. In two, mothers turned down a diet excluding more than four foods, symptoms cleared while feeding the child with an extensively hydrolysed formula, whereas challenges with mother's milk induced immediate reactions. Intestinal permeability was altered during provocation tests with mother's milk sampled before maternal diet. Food allergy during breastfeeding may be due to multiple foods and the inefficacy of the sole CMP elimination in mothers does not rule out food sensitization.     

Sensitization to cow's milk proteins during refeeding of guinea pigs recovering from polydeficient malnutrition

We have previously shown that milk sensitization aggravates intestinal dysfunction in the malnourished guinea pigs, suggesting that it may also impair the recovery from malnutrition. To test this hypothesis, the growing guinea pigs were malnourished by feeding only maize for 7 d and then were refed for 21 d with a balanced diet containing either intact or hydrolyzed cow's milk proteins. The control animals received the hydrolyzed milk protein diet for 28 d. After an initial period of total inhibition of growth owing to maize, guinea pigs gained weight regularly, with both balanced diets, and there was no evidence of mucosal damage at the end of the refeeding period. However, refeeding with intact milk proteins induced milk sensitization, which was demonstrated on the systemic level by the presence of anti-beta-lactoglobulin IgG1 antibodies, and on the local level by the intestinal anaphylaxis measured by the increase in short circuit current induced by beta-lactoglobulin (16.4 +/- 2.6 microA/cm2) in jejunal segments mounted in Ussing chambers. Such an immune sensitization was associated with impaired intestinal permeability, as both the ionic conductance (21.0 +/- 1.6 versus 14.6 +/- 0.7 mS/cm2) and the transepithelial fluxes of horseradish peroxidase (537 +/- 203 versus 152 +/- 28 ng/h x cm2) were significantly increased in guinea pigs refed with the intact milk proteins compared with controls. In contrast, there was no difference in intestinal permeability between controls and guinea pigs refed with the hydrolyzed milk protein diet. These data show that sensitization to cow's milk proteins can develop in guinea pigs recovering from severe malnutrition and may impair full intestinal repair.     

Evaluation of an extensively hydrolyzed casein-whey protein formula in immediate cow's milk protein hypersensitivity

BACKGROUND: Hydrolysate formulas are safe for most infants who are allergic to cow's milk. The purpose of this study was to assess the clinical tolerance and safety of an extensively hydrolyzed casein-whey protein formula in a homogeneous group of patients with immediate cow's milk hypersensitivity. METHODS: The study population consisted of 33 infants in whom immediate cow's milk protein allergy had been diagnosed by clinical data, evidence of specific immunoglobulin E antibodies, and positive results in a controlled challenge test with cow's milk proteins. RESULTS: The hydrolysate was well tolerated in 31 of the 33 patients, which represented a 94% effectiveness. Using 95% confidence intervals generated for a binomial outcome (reaction-no reaction), the casein-whey protein formula is tolerated by 84.4% to 99% of infants with immediate immunoglobulinE-mediated cow's milk protein allergy. These figures are in the range of values recommended for this type of product. CONCLUSIONS: This extensively hydrolyzed casein-whey protein formula is generally safe to feed children with immediate hypersensitivity to cow's milk. However. it is advisable that the first intakes be given under direct medical supervision, in that the occurrence of adverse reactions in highly sensitive infants cannot be unequivocally excluded.     

The role of T lymphocytes in patients with food-sensitive atopic dermatitis

The role of T lymphocytes was assessed in patients with food-sensitive atopic dermatitis (AD). T lymphocytes plus monocytes responded well to ovalbumin or bovine serum albumin (BSA) in children with AD who were sensitive to hen's egg or cow's milk compared with healthy children and children with immediate allergic symptoms who are sensitive to hen's egg or cow's milk. The responding cells were shown to be predominantly CD4+ T lymphocytes. Interleukin-2 activity and interferon-gamma concentrations in culture supernatants of ovalbumin-stimulated peripheral blood mononuclear cells (PBMCs) from patients with AD who were sensitive to hen's egg were significantly higher than those of healthy children and patients sensitive to hen's egg with immediate symptoms. Expression of Fc epsilon R II on B lymphocytes in cultures of ovalbumin-stimulated PBMCs from patients with AD was significantly higher than that of healthy children, but it tended to be lower than that of patients with immediate symptoms. These results suggest that, in patients with AD who are food sensitive, CD4+ T lymphocytes stimulated by food antigens secrete lymphokines such as interleukin-2 and interferon-gamma that are secreted from TH1 clones in mice, and express Fc epsilon R II on B lymphocyte that is induced by interleukin-4 secreted from TH2 clones in mice. Taken together, cell-mediated immunity may also occur in addition to IgE-mediated hypersensitivity in patients with food-sensitive AD.     

Cow's milk intolerance with melena

The clinical features, the results of gastric secretory function tests, and the duodenojejunal morphology of six infants (aged 0.42-1.23 years) with anemia and melena considered to be due to latent cow's milk intolerance (LCMI) were compared with the findings in nine infants (aged 0.19-0.87 years) with cow's milk-induced malabsorption (CMI). The infants with LCMI had a short period of breast feeding, normal weight gain without symptoms of malabsorption, and no atopic history. The maximal acid secretion was decreased (p < 0.01) and the concentration of fasting serum gastrin raised (p < 0.01) compared with the controls. Gastric biopsy revealed epithelial degeneration in three and erosion in one out of four samples. The duodenojejunal biopsy revealed slight changes in two samples, the others being normal. The number of eosinophils was increased in four out of six biopsies. Although the number of intraepithelial lymphocytes was increased in LCMI the rise was not as significant as in children with CMI (p < 0.05). We conclude from our results that LCMI seems to be a seperate clinical entity. The determination of fasting serum gastrin, maximal gastric acid secretion and intraepithelial lymphocytes on duodenojejunal biopsy appear to be helpful in making the diagnosis.     

In vitro diagnosis of cow's milk protein sensitive enteropathy by organ culture method

The criteria that are used at present to diagnose cow's milk protein sensitive enteropathy (CMPSE) are based on an in vivo milk challenge which can be hazardous and life threatening. We have used an organ culture model to determine the usefulness of this technique in establishing the diagnosis of CMPSE on the basis of a single biopsy with in vitro milk challenge. Fourteen infants with diarrhoea clinically suspected to have CMPSE were studied prospectively. On the basis of milk challenge studies seven infants had CMPSE. They had clinical reaction to cow's milk with associated histological changes and depression of alkaline phosphatase levels in the jejunal mucosa. In all seven cases parallel changes in alkaline phosphatase levels were noted in the organ culture specimens of initial biopsy subjected to in vitro challenge. The seven control infants tolerated cow's milk and did not have histological changes. The alkaline phosphatase levels were moderately increased in the jejunal mucosa in five of the seven infants. The alkaline phosphatase levels in the organ culture specimens of initial biopsy were increased after in vitro challenge in all seven infants. This study suggests that organ culture methods may be useful in the vitro diagnosis of CMPSE, and also obviate the need for in vivo oral milk challenges and repeated biopsies.     

Gliadin-specific and cow''s milk protein-specific IgA in human milk

The presence of gliadin-specific IgA and IgG antibodies in colostrum and serum of 140 newly delivered mothers was assessed by enzyme-linked immunosorbent assay. In addition, cow's milk protein (CMP)-specific IgA was determined in the colostrum samples. From 14 of the mothers longitudinal milk samples were obtained after 1 and 2 months of lactation and from 12 mothers after 3 months. Gliadin-specific IgA was found in 97.1% and gliadin-specific IgG in 9.3% of the colostrum samples. Gliadin-specific IgA was detected in mature samples but at significantly lower levels after 1, 2, and 3 months of lactation (p less than 0.01) as compared with colostrum. Gliadin-specific IgA was found in 2.8% of the serum samples and gliadin-specific IgG in 40%; however, the levels of both isotypes were low. CMP-specific IgA was found in 78.1% of the colostrum samples. It is concluded that IgA antibodies to two common food proteins are frequently found in human milk and that food-specific IgA present in milk may play a role in adapting the infant's immune reactions to food antigens in the gut.     

Myths and facts about breastfeeding: does it prevent later atopic disease?

Exclusive human milk feeding during the first 6 months of life, with delayed introduction of solids, is the recommended feeding for human infants. Human milk reduces the incidence and morbidity related to infection and allergy to cow's milk proteins. Dietary maternal restrictions during (late) pregnancy or lactation cannot be recommended, but may be advised in special cases. A maternal elimination diet seems more effective if associated with environmental hypoallergenic intervention(s). Milk from mothers consuming cow's milk proteins contains small amounts of beta-lactoglobulin, which appear to introduce in the majority of infants both atopic and non-atopic tolerance rather than sensitization. However, it is uncertain whether breastfeeding also reduces the incidence of later atopic disease, since its aetiology is multifactorial.     

Iron sufficiency in breast-fed infants and the availability of iron from human milk

Four infants were studied who had been exclusively breast-fed for periods varying from 8 to 18 months. All had grown sufficiently to have exhausted their prenatally acquired iron endowment with respect to meeting current needs for maintaining normal hemoglobin levels. All infants had normal hemoglobin values and normal serum iron values. Studies of iron absorption from breast milk and cow's milk were performed in ten normal adults. The absorption of iron from the human milk was significantly higher. These findings suggest that the iron present in human milk is sufficient to meet the iron requirements of the exclusively breast-fed infant until he approximately triples his birthweight.     

Intolerance of cow's milk and chronic constipation in children

BACKGROUND: Chronic diarrhea is the most common gastrointestinal symptom of intolerance of cow's milk among children. On the basis of a prior open study, we hypothesized that intolerance of cow's milk can also cause severe perianal lesions with pain on defecation and consequent constipation in young children. METHODS: We performed a double-blind, crossover study comparing cow's milk with soy milk in 65 children (age range, 11 to 72 months) with chronic constipation (defined as having one bowel movement every 3 to 15 days). All had been referred to a pediatric gastroenterology clinic and had previously been treated with laxatives without success; 49 had anal fissures and perianal erythema or edema. After 15 days of observation, the patients received cow's milk or soy milk for two weeks. After a one-week washout period, the feedings were reversed. A response was defined as eight or more bowel movements during a treatment period. RESULTS: Forty-four of the 65 children (68 percent) had a response while receiving soy milk. Anal fissures and pain with defecation resolved. None of the children who received cow's milk had a response. In all 44 children with a response, the response was confirmed with a double-blind challenge with cow's milk. Children with a response had a higher frequency of coexistent rhinitis, dermatitis, or bronchospasm than those with no response (11 of 44 children vs. 1 of 21, P=0.05); they were also more likely to have anal fissures and erythema or edema at base line (40 of 44 vs. 9 of 21, P<0.001), evidence of inflammation of the rectal mucosa on biopsy (26 of 44 vs. 5 of 21, P=0.008), and signs of hypersensitivity, such as specific IgE antibodies to cow's-milk antigens (31 of 44 vs. 4 of 21, P<0.001). CONCLUSIONS: In young children, chronic constipation can be a manifestation of intolerance of cow's milk.     

Food allergy: recent findings

Food allergy induces in infancy and childhood a large variety of symptoms which may be trivial in many children, chronic and severe in others and even fatal in rare cases. According to double-blind placebo controlled oral food challenges, cow's milk, egg, wheat and fish are the most common offending foods. Elimination of the offending food(s) is imperative for the management of children with food allergy. An appropriate formula without cow's milk proteins and allergenic epitopes should be given to infants with cow's milk allergy. Breast feeding and selected weaning after the sixth month of life are recommended for the prevention of food allergy in atopic prone babies.     

Mucosal enterokinase activity in cow's milk protein sensitive enteropathy

Enterokinase has a critical role in initiating proteolytic digestion by hydrolysing the conversion of pancreatic trypsinogen into trypsin. The enzyme is synthesised by enterocytes of the proximal small intestine and initially incorporated into the brush border from where it is released into the intestinal lumen by the action of pancreatic secretions. The aim of the study was to analyse enterokinase activity in the duodenal mucosa of infants with diarrhoeal disease including cow's milk protein-sensitive enteropathy. Our observations show that the mean depletion of enterokinase was only 17% compared to 60-80% for other brush border enzymes like disaccharidases, peptidases and alkaline phosphatases in infants with diarrhoea. This suggests that enterokinase activity in the small bowel enteropathies may be dependent not only on the degree of mucosal damage specifically but also on the extent of damage to the goblet cell population where the enzyme is synthesised. Thus the enterokinase activity was reduced in acute and chronic diarrhoea with marked mucosal damage where significant reduction of goblet cell population was evident but the enzyme was relatively little affected when the mucosa was damaged mildly.     

On cyclins, oocytes, and eggs

We studied the preventive effect against allergies in infants who and whose mothers consumed hypoallergenic formulas until 6 months after birth. Mother and infant pairs were divided into three groups, and the infants were monitored for the development of allergies for the first 2 years. In the MD group (n = 102; n = number of infants), the mothers were given a hypoallergenic formula for mothers (MOM HA), which contained hydrolyzed whey protein as the only protein source, as a substitution for cow's milk during late pregnancy and lactation. In the CD group (n = 127), the mothers were given cow's milk during the corresponding period. All infants in the MD and CD groups were exclusively breast-fed or mixed-fed with breast milk and hypoallergenic infant formula (NAN HA), which contains the same hydrolyzed protein as MOM HA. In the AF group (n = 54), the mothers consumed MOM HA and their infants were mixed-fed with breast milk and a cow's milk-based adopted infant formula during the corresponding period. In the MD group, no infants were positive to cow's milk-specific immunoglobulin E (RAST) at 4 months of age, in contrast to 6% and 3% of infants in the CD and AF groups, respectively. The infants in the MD group showed low incidence of various allergies, especially of eczema, as compared to the CD and AF groups. These results suggest that consumption of cow's milk by mothers and cow's milk-based formula feeding to infants elevate the risk of allergies in infants, and that consumption of hypoallergenic formula for pregnant and lactating women and for infants could be helpful in preventing allergy development in infants.     

Breast milk and the prevention of neonatal and preterm gastrointestinal disease states: a new perspective

In this review of the protective properties of human breast milk, a new perspective is taken to underscore the passive and active protection properties of breast milk in providing specific protection against selective gastrointestinal disease affecting the neonate and preterm infant. The normal protective properties of the gastrointestinal epithelial barrier (immunologic and nonimmunologic) are considered as is the development of barriers to antigen absorption in the immature infant human intestine as a background for considering three accelerated gastrointestinal diseases-necrotizing enterocolitis, intestinal allergy, and bacterial gastroenteritis. In each of these conditions, the developmental protective defect is considered and the role of breast milk plays in filling the protective void discussed. Besides considering passive protection of breast milk including the new roles assigned to nutrients such as lactoferrin and nucleotides, and importance of active substances in breast milk such as growth factors, cytokines and hormones are discussed in the context of actively stimulating the infant's own intestinal defenses to function as a protective barrier. Future studies at the cellular and molecular level should be helpful in designing both preventative and treatment strategies to deal with these diseases.     

Production of exfoliative toxin A by Staphylococcus aureus isolated from mastitic cow's milk and farm bulk milk

The production of exfoliative toxins A and B (ETA and ETB) by Staphylococcus aureus isolated from mastitic cow's milk and farm bulk milk was examined by the reverse passive latex agglutination method (RPLA). ETA was detected in 2 (1.2%) of 162 isolates from mastitic cow's milk and in 1 (0.6%) of 166 isolates from farm bulk milk. RPLA titers of these isolates were much lower than in human isolates. No ETB was detected in any of the isolates tested. These ETA-positive isolates belonged to bovine ecovar. They were non-typable using the international phage set for human strains. When these ETA-positive isolates were subcutaneously inoculated into neonatal mice, general exfoliation of the epidermis accompanied by the so-called Nikolsky sign was not recognized. By the immunoblotting and PCR methods, however, ETA and eta gene were recognized in the ETA-positive isolates from mastitic cow's milk and farm bulk milk. These data suggest that ETA is also produced by bovine isolates of S. aureus, but in smaller quantities.     

Allergy to the heat-labile proteins alpha-lactalbumin and beta-lactoglobulin in mare's milk

BACKGROUND: Allergy to mare's milk is rare. Recently, however, mare's milk has been recommended for treatment of various ailments by practitioners of "alternative medicine," and it is available in health food stores. OBJECTIVE: We report a case of allergic reaction to mare's milk in a 51-year-old woman who was able to tolerate cow's milk. METHODS: The protein composition of mare's milk was determined by methods based on measurement of nitrogen content. The patient underwent prick and intracutaneous tests with commercially available bovine milk proteins and several mare's milk preparations, including mare's milk granulate and boiled mare's milk. RAST and immunoblotting were also performed. RESULTS: Results of skin testing and RAST with cow's milk were negative but demonstrated an IgE-mediated allergy to mare's milk. Immunoblotting revealed two allergen bands with molecular weights of 16 and 18 kd, most likely representing the whey proteins alpha-lactalbumin and beta-lactoglobulin. The bands disappeared after the mare's milk was boiled, indicating that the proteins are heat-labile. CONCLUSION: The results of this study demonstrate the existence of an IgE-mediated mare's milk allergy caused by low molecular weight heat-labile proteins, most likely alpha-lactalbumin and beta-lactoglobulin, which do not cross-react with the corresponding whey proteins in cow's milk.     

Tumor necrosis factor alpha regulates proliferation in a mouse intestinal cell line

BACKGROUND & AIMS: Tumor necrosis factor (TNF)-alpha is a prominent cytokine in the pathogenesis of inflammatory bowel disease, yet its effects on the intestinal epithelium remain poorly understood. This study was designed to investigate the action of TNF-alpha on intestinal cell proliferation. METHODS: Young adult mouse colon cells were studied under nontransformed conditions with epidermal growth factor, keratinocyte growth factor, insulin-like growth factor 1, or serum in the presence or absence of TNF-alpha and cell numbers determined. The expression and independent actions of the 55-kilodalton TNF-alpha R1 and 75-kilodalton TNF-alpha R2 receptors were studied by immunologic methods. RESULTS: TNF-alpha stimulated proliferation at 0.1 and 1 ng/mL and inhibited proliferation at 100 and 1000 ng/mL without altering cell viability. TNF-alpha inhibited the mitogenic effect of growth factors and epidermal growth factor receptor tyrosine phosphorylation. TNF-alpha R1 receptor agonist antibody inhibited proliferation, whereas a TNF-alpha R2 receptor-blocking antibody prevented the proliferative effect of low-dose TNF-alpha. CONCLUSIONS: TNF-alpha displays a novel influence on intestinal cell growth, stimulating proliferation at physiological concentrations and inhibiting proliferation at pathological concentrations. The regulation of intestinal cell mitogenesis by TNF-alpha seems to be mediated differentially by the two TNF-alpha receptors, with the TNF-alpha R1 receptor inhibiting proliferation and the TNF-alpha R2 receptor promoting proliferation.