Allergenicity of mare's milk in children with cow's milk allergy

BACKGROUND: Cow's milk allergy is a common disease of infancy and early childhood. If the baby is not breast-fed, a substitute for cow's milk formula is necessary. OBJECTIVE: The aim of this study was to investigate, in vitro and in vivo, the allergenicity of mare's milk in a population of selected children with severe IgE-mediated cow's milk allergy. METHODS: Twenty-five children (17 male and 8 female) aged 19 to 72 months (median age 34 months) with IgE-mediated cow's milk allergy were selected for this study. All the children underwent skin prick tests with cow's milk and mare's milk and double-blind placebo-controlled oral food challenge (DBPCOFC) with fresh cow's milk, fresh mare's milk, and, as placebo, a soy formula (Isomil, Abbott, Campoverde, Italy). We performed immunoblotting of cow's and mare's milk developed with IgE from allergic children. RESULTS: All the children showed strong positive skin test responses to cow's milk (4+); 2 children had positive skin test responses to mare's milk (2+). All children had positive DBPCOFCs to cow's milk; one child had a positive DBPCOFC to mare's milk. No children reacted to the placebo (Isomil). In the cow's milk, some proteins are able to strongly react with human IgE; when the sera are tested with mare's milk, the bands corresponding to the same proteins are recognized by a lower percentage of sera. CONCLUSION: These data suggest that mare's milk can be regarded as a good substitute of cow's milk in most children with severe IgE-mediated cow's milk allergy. It would be prudent, however, to confirm its tolerability by a supervised titrated oral challenge test.     

Skin testing with ultra-heat-treated (UHT) cow's milk in children with cow's milk allergy

BackgroundA large proportion of cow's milk (CM)-allergic children are able to tolerate extensively heated forms of CM such as baked goods. Little is known about whether ultra-heat-treated (UHT) forms of cow's milk are immunologically similar to extensively heated cow's milk and therefore may be tolerated by these children.ObjectiveTo determine whether skin test wheal size using UHT CM was significantly different from other forms of CM and CM extracts.MethodsChildren presenting for oral food challenges with either extensively heated or unheated cow's milk underwent skin prick test (SPT) to commercial CM, UHT CM, evaporated CM, and fresh whole CM. The results were compared between groups of children.ResultsAt study exit, only 14% of children were avoiding all forms of CM, compared with 70% at study entry. No difference was seen in the mean SPT results for UHT CM between those children that could tolerate heated CM compared with those that could not. The mean SPT result for casein was significantly lower in those that could tolerate heated CM. However, within the group of heated milk–tolerant children, the mean SPT for UHT CM was significantly lower than the SPT for fresh whole CM.ConclusionUltra-heat-treated CM does not behave significantly differently from other forms of CM when evaluated by SPT in heated milk–allergic vs heated milk–tolerant children. This suggests that UHT CM is not sufficiently immunologically different from unheated CM to be tolerated by heated CM–tolerant children.     

Prenatal allergen contact with milk proteins

Background Cellular proliferation to various allergens (Dermatophagoides pleronyssinus, β-lactoglobulin, bovine serum albumin, ovalbumin) has been found in cord blood cells. Whether this reflects a sensitization during foetal life is uncertain. Objective We studied the cellular reactivity and cytokine production of cord blood cells in response to cow's milk proteins in a randomely selected group of newborns. The delineation of possible in utero allergen contact was attempted. Objective Cord blood mononuclear cells from 39 neonates were incubated with cow's milk proteins (α-lactalbumin, β-lactoglobulin, casein, α-casein, β-casein, k-casein, bovine serum albumin) for 7 days, and proliferation was assessed by incorporation of [³H]thymidine. Cord blood cell-derived interferon-γ (IFNγ) and interleukin-4 (IL-4) secretion was evaluated in response to allergen or phytohaemagglutinin (PHA) stimulation. Results A pronounced proliferation of cells stimulated with α-lactalbumin (ALA; mean stimulation index 8.0, 95% confidence interval 5.2–10.8), β-lactoglobulin (BLG; mean stimulation index 5.9, 95% confidence interval 3.2–8.6) and α-casein (2.6, 95% confidence interval 2.9–9.1), as opposed to unstimulated cells in medium, was found. No correlation was found between cellular proliferation to milk proteins and parental atopy, maternal total IgE or cord blood IgE. IFNγ production (but not IL-4) was inducible by PHA (range 429–1810 pg/ml), but only in one individual upon stimulation with BLG. Preferentially, reduced IFNγ levels were found in individuals with positive parental allergic history. Conclusion The recognition of allergen by cord blood cells indicates that allergen priming must occur prenatally. The relevance for subsequent sensitization is unclear.     

Microbiological study of breast milk with special reference to its storage in milk bank

Sixty five breast milk samples were subjected to bacteriological studies; each sample thrice (in fresh state, after heating at 1000C and freezing for 5 days at -20 degrees C). In fresh State, Staphylococci and diphtheroids were predominant organisms. After heating, none of the samples showed any micro-organisms while after freezing for 5 days, all the samples showing presence of micro-organisms earlier, showed decrease in colony counts. None of the milk samples showed inclusions of cytomegalovirus. Serum samples of mothers tested for Hepatitis B surface antigen and human immunodeficiency virus antibodies gave negative results. Such studies i.e. screening of breast milk samples are important if human milk is to be stored in banks and supplied to premature (high risk group) children.     

IgE antibodies to hydrolysates of cow milk proteins in children with cow milk allergy

Milk substitutes such as protein hydrolysates are largely used in children with cow milk allergy. The clinical benefit of these preparations is still a matter of debate. In the present study, IgE directed against protein hydrolysates was detected in 6/13 in children with cow milk allergy.     

Hypervitaminosis D associated with drinking milk.

BACKGROUND. Vitamin D has been added to milk in the United States since the 1930s to prevent rickets. We report the unusual occurrence of eight cases of vitamin D intoxication that appear to have been caused by excessive vitamin D fortification of dairy milk. METHODS. Medical records were reviewed and a dietary questionnaire was sent to eight patients who had unexplained hypervitaminosis D. Vitamin D analyses with high-performance liquid chromatography were performed on samples of the patients' serum, the dairy milk they drank, and the vitamin D concentrate added to the milk. RESULTS. All eight patients drank milk produced by a local dairy in amounts ranging from 1/2 to 3 cups (118 to 710 ml) daily. All had elevated serum 25-hydroxyvitamin D concentrations (mean [+/- SD], 731 +/- 434 nmol per liter [293 +/- 174 ng per milliliter]). Six of the eight patients had elevated serum vitamin D3 concentrations. Of the eight patients, seven had hypercalcemia and one had hypercalciuria but normocalcemia (mean serum calcium, 3.14 +/- 0.51 mmol per liter [12.6 +/- 2.1 mg per deciliter]). Analysis of the dairy's vitamin D-fortified milk revealed concentrations of vitamin D3 (cholecalciferol) that ranged from undetectable to as high as 232,565 IU per quart (245,840 IU per liter). An analysis of the concentrate that was used to fortify the milk, labeled as containing vitamin D2 (ergocalciferol), revealed that it contained vitamin D3. CONCLUSIONS. Hypervitaminosis D may result from drinking milk that is incorrectly and excessively fortified with vitamin D. Milk that is fortified with vitamin D must be carefully monitored.     

Human milk antibodies with polysaccharide kinase activity

It was shown for the first time that a small fraction of milk secretory IgA (sIgA) is tightly bound to oligosaccharides (oligoSACs) and polysaccharides (polySACs). The ability of sIgA to phosphorylate oligo- and polysaccharides was shown to be an intrinsic property of this antibody. In contrast to known kinases, sIgAs with polysaccharide kinase activity can transfer phosphoryl group to oligo- and polysaccharides not only from [gamma-(32)P]ATP but can also use [(32)P]orthophosphate as a substrate of phosphorylation reaction. An extremely unusual property of polysaccharide kinase Abs is their high affinity for orthophosphate (K(m) = 15-77 microM), and orthophosphate is a better substrate than ATP. Two first examples of natural abzymes (Abzs) with synthetic activity were milk sIgA with protein and lipid kinase activities. Polysaccharide kinase sIgA of human milk is the third example of natural antibodies (Abs) with synthetic activity.     

Immunoglobulin E and immunoglobulin G4 antibodies to cow''s milk in children with cow''s milk allergy

The presence of cow's milk specific antibodies of immunoglobulin E and G4 classes were studied in 47 children with a positive clinical history of cow's milk allergy. The children were challenged with cow's milk orally. The clinical diagnosis was verified by immediate reactions in 25 patients while 22 had late reactions or were provocation test negative in spite of the clinical history. There was no relation between levels of cow's milk specific IgG4 antibodies and provocation test result, i.e. neither with immediate or late reactions. Total IgF. Was elevated above + 1 SD for age in 31 41 tested patients. Of these, 29 had immediate type reactions to cow's milk wheat flour and/or egg white, while only two of 10 children with IgE of less than +1 SD had a demonstrable allergy to any of these foods. The sensitivity was 80%. Specific IgE antibodies to cow's milk were demonstrated in 11 of 14 children with immediate reactions and in three of 15 who were provocation test negative or had only late reactions. This means a sensitivity of 79% and a specificity of 80%. At least had only late reactions. This means a sensitivity of 79% and a specificity of 80%. At least one of the four patients with specific IgE but negative provocation test result had earlier shown an immediate reaction when challenged with cow's milk indicating that the specific IgE antibodies were not truly "false" positive reactions but a consequence of previous allergy. Our results confirm an association between elevated total serum IgE and food allergy and an association between positive RAST to cow's milk and positive provocations in young children. We did not find any evidence for specific IgE-1 antibodies playing a role in these patients.     

Enterococci in milk and milk products.

Enterococci are widely distributed in nature. They gain entry into milk and milk products through the water supply, equipment, and insanitary and unhygienic conditions of production and handling. They have been incriminated as direct or indirect agents of disease. The evidence concerning their involvement is only circumstantial. These reports are also disputed as the disease symptoms have not been experimentally induced in animal models. However, there is sufficient evidence to indicate that prolific growth of enterococci in foods may lead to formation of clinically significant levels of pressor amines. These amines are very thermostable and therefore remain active even after heat processing, which eliminates all viable streptococci. These pressor amines may be involved in the onset of migraine attacks and produce hypersensitive crises in psychiatric patients who are being treated with monoamine oxidase inhibitors for depression.     

Allergenicity of goat’s milk in children with cow’s milk allergy

BACKGROUND: Cow's milk allergy (CMA) is a common disease of infancy and childhood. An appropriate cow's milk (CM) substitute is necessary for feeding babies with CMA. CM substitutes are soy formulas and casein- or whey-based extensively hydrolyzed formulas. In several countries, including Italy, goat's milk (GM) formulas are available, and some physicians recommend them for feeding babies with CMA. OBJECTIVE: We sought to investigate, in vitro and in vivo, the allergenicity of GM in 26 children with proven IgE-mediated CMA. METHODS: All the children underwent skin tests with CM and GM; detection of specific serum IgE to CM and GM; and double-blind, placebo-controlled, oral food challenges (DBPCOFCs) with fresh CM, GM, and, as placebo, a soy formula (Isomil, Abbott, Italy). CAP inhibition and immunoblotting inhibition assays were also carried out in 1 of 26 and 4 of 26 children with positive RAST results to both CM and GM, respectively. RESULTS: All the children had positive skin test responses and CAP results to both CM and GM, all had positive DBPCOFC results to CM, and 24 of 26 had positive DBPCOFCs to GM. In CAP inhibition tests, preincubation of serum with CM or GM strongly inhibited IgE either to CM or to GM. In immunoblotting inhibition assays, preincubation with CM completely extinguished reactivity to GM, whereas GM partially inhibited reactivity to CM. CONCLUSIONS: These data strongly indicate that GM is not an appropriate CM substitute for children with IgE-mediated CMA. A warning on the lack of safety of GM for children with CMA should be on the label of GM formulas to prevent severe allergic reactions in babies with CMA.     

Comparison of intestinal anaphylactic reactions in sensitized mice challenged with untreated bovine milk and homogenized bovine milk

Outbred NMRI mice were sensitized for high IgE production either by subcutaneous injections of low doses of untreated bovine milk or homogenized bovine milk in combination with intraperitoneal injections of Freund's Complete Adjuvant or by oral administration of untreated or homogenized bovine milk without adjuvant. When analysed in murine passive cutaneous anaphylaxis test both types of milk resulted in production of reaginic antibodies against bovine milk proteins when given subcutaneously. When given orally, homogenized milk resulted in reagin production in 10 out of 14 mice, whereas untreated milk resulted in reagin production in only one out of 12 mice. The sensitized mice, and control mice, were orally challenged with either untreated milk, homogenized milk or 0.9% NaCl. Examination of the intestines 40 min after oral administration revealed that homogenized milk, contrary to untreated milk or 0.9% NaCl, resulted in a large increase in the mass of the proximal gut segment of mice sensitized orally with homogenized milk compared with control mice orally challenged with saline (P less than 0.001), and only mice both sensitized and challenged orally with homogenized milk showed degranulation of mast cells in the intestinal wall. By contrast, subcutaneously sensitized mice or mice sensitized orally with untreated bovine milk showed no significant intestinal reaction upon oral challenge with either homogenized or untreated bovine milk. These observations may indicate that the route of sensitization is of great importance when intestinal reactions are to be studied, and that homogenization of bovine milk may render the milk more aggressive with respect to its ability to induce intestinal reactions. The study indicates that mice may be an attractive experimental animal model for mimicking the intestinal anaphylactic reactions of cow milk-allergic humans.     

Passage of cows' milk protein in breast milk

An enzyme-linked immunosorbent assay has been used to search for lactoglobulin and bovine casein in breast milk. Initial results suggest this may be a simple and useful method for such studies. Detection and prevention of antigen passage could influence the prevalence of atopic disease.     

Correlation of lymphocyte proliferating cell nuclear antigen expression with dietary cow's milk antigen load in infants with allergy to cow's milk.

BACKGROUND: Controversial results have been reported on the participation and diagnostic value of lymphocyte reactivity in cow's milk (CM) allergy. In this study, we used a specific nuclear marker to evaluate lymphocyte proliferation in IgE-mediated CM allergy in infants, and examine its relation with diets containing different CM antigen loads. METHODS: Infants with IgE-mediated CM allergy, as assessed by open provocation and RAST, were grouped according to their exclusive diet, either CM formulae, breast feeding, or hydrolysed whey formulae. A group of non-atopic infants receiving CM was also examined. Lymphocyte proliferation to beta-lactoglobulin was evaluated by quantitation of the proliferating cell nuclear antigen (PCNA) expression in peripheral blood mononuclear cells, by flow cytometry. Immunophenotypic surface markers were also examined. RESULTS: A marked difference of PCNA expression between CM-fed allergic infants and healthy controls was observed (p<0.001). In this setting, PCNA expression >/=10% was highly specific and sensitive as a marker of CM allergy in CM-fed infants. Moreover, a significant correlation (p<0.001) between antigen load and PCNA was established in CM-allergic infants under different diets, higher values obtained with increasing antigen loads. In addition, within the group fed hydrolyzed formulae, low-molecular-weight products resulted in marginally lower PCNA expression than higher-molecular-weight formulae. No differences in immunophenotype were found, with the exception of a higher CD23 expression in the breast-fed group. CONCLUSIONS: PCNA could be a useful marker in the assessment of lymphocyte proliferation to CM antigens. Low CM antigen diets are related with reduced lymphocyte reactivity, which may partly explain the clinical benefit observed with such diets.     

Serum antibodies against native, processed and digested cow's milk proteins in children with cow's milk protein intolerance

Children with or without cow's milk protein intolerance were investigated for serum antibodies to native cow's milk proteins, processed cow's milk proteins and native- and pepsin-digested β-lactoglobulin. Antibodies of IgG and IgA isotypes were determined with ELISA and those of IgE isotype with RAST. Children who exhibited slowly appearing untoward reactions to cow's milk feeding had significantly higher titres of IgG antibodies against both native and digested β-lactoglobulin than children with an immediate type of reaction or no intolerance. The IgG antibodies to pepsin-digested β-lactoglobulin were efficiently inhibited by native β-lactoglobulin even at low concentrations, which suggests that there were no antibodies specific for the degraded proteins. The children with slow reactions to cow's milk also tended to have higher antibody levels of the IgG and IgA isotypes to both native and processed cow's milk. Antibodies to this mix of antigens discriminated less well, however, than the antibodies to isolated β-lactoglobulin between the groups of children with slowly appearing reactions, with immediate reactions and the controls. Seven out of nine children with an immediate type of reaction to cow's milk protein had IgE antibodies against cow's milk protein determined with the RAST, while this type of antibodies could not be demonstrated in any child in the two other groups. Using processed or digested protein as antigen did not increase the sensitivity of the antibody determinations.