Bovine β-Lactoglobulin in the Human Milk

ABSTRACT. Human milk samples ( n =232) collected during the whole lactation period from 25 healthy, Swedish mothers were analyzed by radioimmunologic method for content of bovine β-lacto-globulin. Detectable amounts (5-800 μ/1) were found in 93 of 232 milk samples (40%). Six mothers had no detectable β-lactoglobulin in their breast milk on any occasion. Two mothers had measurable /Mactoglobulin in all their milk samples. No correlation was found between daily cow's milk intake and concentration of β-lactoglobulin in the milk samples. Six mothers with allergic symptoms such as asthma, hay-fever, eczema all had detectable amounts of β-lactoglobulin in their milk. Of 19 mothers without allergy, 13 had detectable amounts. This difference did not show statistical significance. The presence of symptoms in the infant such as diarrhoea, vomiting, colic, exanthema was significantly correlated to high levels of β-lactoglobulin in the milk. Bovine β-lactoglobulin was also detected in 7 of 13 serum samples. The two mothers with detectable β-lactoglobulin in all milk samples had the highest serum values, and their infants suffered from gastro-intestinal symptoms, weight decline and exanthema.     

Fetal peripheral blood mononuclear cell proliferative responses to mitogenic and allergenic stimuli during gestation

Blood samples were obtained from fetuses and premature babies (n=51) (15-34 weeks gestation) to determine at what stage the fetal immune system was able to produce a positive proliferative response to common allergens. Peripheral blood mononuclear cells (PB MC) were stimulated with the mitogen, phytohaemagglutinin (PHA), and the allergens, house dust mite, cat fur. birch tree pollen, β-lactoglobulin, ovalbumin and bee venom (mellitin). Results were expressed as ratios of stimulated to unstimulated ³H thymidine incorporation, and as percent positive responders. There was an increase in proliferation ratio which correlated with increasing gestational age for PHA (p < 0.0001), cat fur (p=0.042), birch pollen (p=0.022) and β-lactoglobulin (p=0, 006). The point in gestation when cells from some individuals began responding to the allergens with a ratio of 2. 0 was at approximately 22 weeks. PBMC proliferative response ratios were higher from samples from babies > 22 weeks gestation compared to < 22 weeks for the mitogen and all allergens, except mellitin. There was also a greater proportion of positive responders from samples > 22 weeks compared to < 22 weeks for the mitogen and all allergens, except mellitin. Maternal exposure to birch pollen, which has a discrete season, was assessed to determine whether exposure had occurred at 22 weeks gestation or beyond. Results showed a higher prolifera tive response in infant cells stimulated with birch pollen (p=0.005) and higher proportion of positive responders (p=0.01) in the group of babies whose mothers had been exposed to hirch pollen beyond 22 weeks, compared to those whose mothers had not been so exposed. These results suggest that in utero fetal exposure to an allergen from around 22 weeks gestation may result in primary sensitisation to that allergen, leading to positive proliferative responses, at birth.     

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 β-lactoglobulin-specific immunoglobulin A and α-casein-specific immunoglobulin A levels was observed over the first 5 days in all groups. The mean level of β-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.     

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, β-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 β-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 β-cell surface proteins in the aetiology of IDDM.     

Serum Antibodies to Cow's Milk Proteins in Pediatric Inflammatory Bowel Disease

ABSTRACT. Serum antibodies to five cow's milk proteins (α-casein, bovine serum albumin, β-lactoglobu-lin A and B, and α-lactalbumin) were investigated in young patients with inflammatory bowel disease (56 Crohn's disease, 24 ulcerative colitis). IgG antibodies against bovine serum albumin, β-lactoglobulin A and β-lactoglobulin B were higher in Crohn's disease patients than in those with ulcerative colitis or the controls. IgG anti-bovine serum albumin antibodies were higher in those Crohn's disease patients who had higher scores of disease activity. Finally, IgA antibodies to α-casein were higher in patients with Crohn's disease and ulcerative colitis when compared to controls. These findings may be due to increased uptake of dietary antigens or enhanced immunological response occurring in Crohn's disease patients.     

Cow's milk protein intolerance and chronic constipation in children

Cow's milk protein (CMP) allergy was investigated in 25 children (age-range 3 months to 11 years) with chronic constipation. A diagnosis of constipation was made on the basis of a history of painful elimination of hard stools for at least 1 month, whether or not associated with a reduced frequency of stools or soiling. The children were evaluated using clinical parameters and the following laboratory tests: total serum immunoglobulin E (IgE); specific IgE (radioallergosorbent test [RAST]) for whole cow's milk, α-lactoalbumin, β-lactoglobulin, and a food group; and skin-prick tests with whole milk, α-lactoalbumin, β-lactoglobulin, and casein. Following the evaluation, the children were submitted to a CMP-free diet for a period of 4 weeks. In seven patients (28%), constipation disappeared during the CMP-free diet and reappeared within 48–72 h following challenge with cow's milk. In two infants a rectal biopsy revealed allergic colitis and they therefore did not undergo the challenge. High serum levels of total IgE were observed in five of the children who showed a clinical improvement (71%), a positive skin-test in two (29%), and detectable specific IgE in two (29%). These results suggest that CMP allergy or intolerance should be considered as a cause of chronic refractory constipation in children, although the underlying mechanism still require further investigation.     

Breast Feeding Modifies Production of SlgA Cow's Milk-Antibodies in Infants

ABSTRACT. 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-β-lactoglobulin were determined by 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 ≤0.01). The effect of breast-feeding was seen even after nursing period of only 3 weeks. Infants without risk of allergy fed cow's milk exclusively had higher SIgA anti-casein ( p < 0.03) and anti-β-lactoglobulin concentrations than low risk infants at the age of 6 months. These data show modifying effect of breast feeding on salivary SIgA production against cow's milk protein.     

Betaine and homocysteine concentrations in infant formulae and breast milk

BACKGROUND: Early diagnosis and treatment of homocystinuria with methionine-free or low methionine formulae significantly improve prognosis in children found by newborn screening and treated soon after birth. Betaine (Bet) supplementation is also an effective strategy for dietary treatment of homocystinuria. However, previous reports on diet therapy have only examined methionine and cysteine concentrations but not those of Bet and homocysteine (Hcy) in infant diets. METHODS: We measured Bet and Hcy concentrations in three methionine-free formulae, five infant starter formulae, and 33 samples of human breast milk from 10 mothers. RESULTS: In methionine-free formulae, Hcy but not Bet was isolated. However, in human breast milk and infant starter formulae, both Bet and Hcy were detected. However, the Bet concentration was not sufficient for the treatment. CONCLUSIONS: Our study suggests that methionine-free formulae should have Hcy eliminated and be fortified with Bet to the concentration of 150 mg/dL for the treatment of homocystinuria.     

Human milk proteins may interfere in ELISA measurements of bovine β-lactoglobulin in human milk

It is widely believed that cow's milk proteins ingested by the mother, in particular β-lactoglobulin ([β-LG), can pass into breast milk and thus sensitize predisposed infants. However, studies to evaluate bovine β-LG in human milk have given conflicting results. The aim of this study was to analyse the correlation between the amount of cow's milk in the mother's diet and the presence of bovine β-LG in breast milk. Human milk samples from 14 healthy non-atopic women on diets with different cow's milk contents were examined. The total concentration of bovine β-LG or β-LG immuno-like proteins (β-LGIP) was determined by enzyme-linked immunosorbent assay (ELISA). Two separation procedures utilizing ELISA plates and an affinity chromatography column were set up to identify the human whey components recognized by the anti-β-LG antibodies. β-LGIP reactivities of milk from three groups on different diets were not significantly different. After splitting the antigen antibody complexes, three main protein components, human lactoferrin, human β-casein and human α-lactalbumin, were identified. This study would suggest that, at least in healthy subjects, false-positive results in ELISA determinations of bovine β-LG in human milk might be due to cross-reactions between polyclonal antibodies and different protein antigens.     

Solubility and digestibility of milk proteins in infant formulas exposed to different heat treatments.

There are presently several fundamentally different technologies to produce infant formulas (IF), such as sterilization, spray-drying, and treatment at ultrahigh temperature (UHT). The effects of heat treatment on milk proteins in IF were analyzed by column chromatography, gel electrophoresis, and Kjeldahl analysis, revealing strong protein-protein and protein-lipid interactions in processed milk. These interactions were more pronounced in conventionally (in-can) sterilized than in spray-dried (powdered) and UHT products confirming their temperature dependency. Analysis of raw materials, intermediate and end products of IF processing revealed that after homogenization the first indications of protein denaturation occurred, but that in-can sterilization as the final heat treatment caused irreversible denaturation of proteins and strong protein-lipid interactions. Lowering of the pH to 4-5, which is physiological for the stomach of young infants, enhanced the interactions. Support for an impairment of protein digestibility was given by in vitro analysis of protein digestibility, demonstrating significantly lower digestibility of in-can sterilized IF compared to their spray-dried and UHT counterparts. To investigate the effect of heat treatment on chemical reactions, i.e., occurrence of Maillard products, we showed by a fluorimetric assay that the amount of "available" lysine is lower in sterilized than in powdered IF. Our findings suggest that a more differentiated view regarding the protein quality of IF is needed.     

Increased frequency of IgM antibodies to cow's milk proteins in Hungarian children with newly diagnosed insulin-dependent diabetes mellitus

We investigated the association between serum antibodies to cow’s milk proteins and insulin-dependent diabetes mellitus (IDDM) in Hungarian children. Forty-eight children 1.0–17.1 years of age with newly diagnosed IDDM and 74 control children 1.0–16.0 years of age were studied for serum IgG, IgA and IgM antibodies to cow’s milk, β-lactoglobulin, bovine serum albumin and ovalbumin by enzyme-linked immunosorbent assays. The specificity of IgM antibodies to β-lactoglobulin and bovine serum albumin was controlled by Western blot. The levels of IgG and IgA antibodies to cow’s milk proteins were similar in children with and without IDDM, with the exception of slightly increased levels of IgA antibodies to β-lactoglobulin in diabetic children (P = 0.05). The levels of IgM antibodies to cow’s milk were significantly higher in IDDM patients than in control children (P = 0.0002). Children with IDDM more often had IgM antibodies to β-lactoglobulin (46.3% vs 18.8%; P = 0.002) and bovine serum albumin (87.8% vs 49.3%, P < 0.0001) than control children. Neither the levels of IgG or IgA antibodies to ovalbumin nor the frequency of IgM antibodies to ovalbumin differed between diabetic and control children. Conclusion In Hungarian children, clinical manifestation of IDDM is often associated with IgM antibody response to cow’s milk protein and its fractions, β-lactoglobulin and bovine serum albumin, indicating a loss of immunological tolerance to these proteins. IgG and IgA antibodies to cow’s milk proteins, associated with an early introduction of cow’s milk in diet, seem to play a minor role in the development of childhood IDDM in Hungary. Received: 14 November 1995 Accepted: 3 March 1996     

Characterization of antigens and allergens in hypo-allergenic infant formulae

The antigenicity and allergenicity of so-called hypo-allergenic infant formulae is mainly determined by the degree of hydrolysis and ultrafiltration. Five different formulae were investigated by means of immunoblotting and RAST in order to characterize the antigens and allergens regarding their molecular weights, molecular origin and their ability to bind human IgG and IgE antibodies: A non hydrolysed infant formula (I-F), a mixture of the major cow's milk proteins (PM), a whey-based infant formula (W-H), a whey-based and ultra-filtrated infant formula (U-H), a casein/whey-based infant formula (CW-H). By immunoblotting we demonstrated that all tested formulae still contain antigens with molecular weights from 3 to 67 kD. But when compared with I-F and PM the antigen content of the hydrolysed formulae was considerably lower. The lowest antigen content could be demonstrated in U-H, which contains casein fragments (3–6 kD) and beta-lactoglobulin and its fragments (6–18 kD). W-H and CW-H contain bovine serum albumin, beta-lactoglobulin, casein and their fragments (3–67 kD). All hydrolysed formulae tested showed a reduced IgE-binding capacity. Three out of 12 cow's milk allergic children possessed IgE binding to U-H or W-H, and 5 of them IgE against CW-H.Conclusion The enzymatic hydrolysis plus ultra-filtration seems to be the most efficient method to reduce the antigen content of so-called hypo-allergenic infant formuale.     

β2-MICROGLOBULIN IN CEREBROSPINAL FLUID FROM CHILDREN WITH DIFFERENT DISEASES

Abstract. Kaas Ibsen, K. (The University Clinic of Paediatrics, Children's Hospital Fuglebakken, Copenhagen, Denmark). β₂-microglobulin in cerebrospinal fluid from children with different diseases. Acta Paediatr Scand, 69: 633, 1980.—β₂-microglobulin concentrations in cerebrospinal fluid (CSF) were measured in a prospective study on 56 children 0–12 years old. In all the patients with virus meningitis values of β₂-microglobulin exceeded 3000 µg/l (=10.825 µg/l). The highest value (48.096 µg/l) of β₂-microglobulin in CSF was found in a 13-day-old infant with serious herpes simplex meningitis. The value was 50 times the values found in normal children. None of the patients with fever of other origin had values exceeding 3500 µg/l, except for one patient with facial nerve paresis and 3 patients with sepsis. Some correlation between the concentrations of β₂-microglobulin and albumin was found in the diagnostic groups as a whole, while this correlation disappeared when considering each patient individually. The significance of β₂-microglobulin as a guide in serious infections is discussed.     

Dietary Bovine β Mactoglobulin is Transferred to Human Milk

ABSTRACT. Human milk from 38 mothers was analysed by radioimmunological method for content of bovine (5-Iactoglobulin. Detectable amounts (5-33 μg/1) of immunoactive β-lactoglobulin were found in 18 human milk samples. Milk from 3 mothers, whose infants suffered from infantile colic contained high amounts of fi-lactoglobulin (32, 18 and 14 μg/1 respectively). With the mothers on a cow's milk free diet the contents fell to non-detectable amounts in two mothers and to 6 μg/1 in the third. All three infants became free from colic,     

Advances in the management of digestive problems during the first months of life

A study was carried out on 168 full-term infants with digestive problems such as regurgitation and/or constipation to evaluate the efficacy of new infant formulas containing partially hydrolysed whey protein, modified vegetable oil with a high β-palmitic acid content, prebiotic oligosaccharides and starch. Infants receiving the new formulas had an increase in stool frequency of 0.60 between day 1 and day 7 (95% CI 0.19–1.01; p =0.004) and 0.53 (95% CI 0.11–0.90; p =0.015) between day 7 and day 14. A reduction of 1.06 in the number of regurgitation episodes was reported between day 1 and day 7 (95% CI 0.24–1.88; p =0.012) and 1.31 (95% CI 0.42–2.21; p =0.005) between day 7 and day 14.
Conclusion: A prebiotic mixture of galacto-/fructo-oligosaccharides with a high β-palmitic acid content may reduce digestive problems and improve intestinal tolerance in infants during the first months of life.     

Cow's milk and ovalbumin-specific IgG and IgA in children with eczema: low β-lactoglobulin-specific IgG4 levels are associated with cow's milk allergy

To cite this article: Savilahti EM, Viljanen M, Kuitunen M, Savilahti E. Cow's milk and ovalbumin-specific IgG and IgA in children with eczema: low β-lactoglobulin-specific IgG4 levels are associated with cow's milk allergy. Pediatric Allergy Immunology 2012: 23: 590-596. ABSTRACT: Tolerance to allergens may partly depend on allergen-specific IgG and IgG subclasses and IgA antibodies. We investigated whether specific IgG and IgG subclasses and IgA antibodies to β-lactoglobulin, α-casein, and ovalbumin differed between infants who had verified cow's milk allergy (CMA) and infants with cow's milk (CM)-associated eczema, but negative CM oral challenge. The study population comprised 95 infants with clinical eczema that was by history associated with the consumption of CM. After an elimination period, a double-blind, placebo-controlled (DBPC) CM oral challenge confirmed CMA in 45 infants. Skin prick tests (SPT) were performed with CM and hen's egg. Serum levels of IgE antibodies to CM and hen's egg were measured with UniCAP (Phadia, Uppsala, Sweden), and levels of IgA, IgG, IgG1, and IgG4 antibodies to β-lactoglobulin, α-casein, and ovalbumin were measured with enzyme-linked immunosorbent assay. We observed that infants with CMA had lower IgG4 levels to β-lactoglobulin than infants with negative DBPC CM challenge (p?=?0.004). Positive CM SPT was associated with lower IgG4 levels to α-casein (p?=?0.04). The relation of CM IgE to β-lactoglobulin and α-casein IgG4 was higher in CMA than in infants with negative challenge (p?相似文献   

Longitudinal monitoring of bone mineral density in thalassemic patients. Genetic structure and osteoporosis

The changes in bone mineral density (BMD) measured by single photon absorptiometry (SPA) using two observations conducted over a period of 2 years were examined in 54 thalassemic subjects [ 30 F(A)and 24 M (B)] with a chronological age ranging from 2.6 to 22.6 years and in 27 sex- and age-matched controls (C). Each category (A. B and C) was divided into three groups according to pubertal signs: pre-pubertal subjects (A1, B1 and C1): peri-pubertal subjects (A2, B2 and C2) and pubertal subjects from the first observation (A3, B3 and C3). Furthermore, each group of patients was divided into sub-groups on the basis of haematological phenotypes, those with a more severe form were called β⁰/β⁰ while those with other forms were called "others". The most significant findings were the following: the presence of a more severe reduction of the bone mineral density in patients with the β⁰/β⁰ phenotype than in patients with the "others" phenotype; patients with hypogonadism corresponded to the β⁰/β⁰ phenotype, while those with spontaneous puberty corresponded to the "others" phenotype. In conclusion, since puberty and the degree of bone mineral density are related to the haematological phenotype, puberty (spontaneous or induced) positively influences the bone mineral density only at the start of puberty, while subsequently, the degree of osteoporosis is the expression of widespread and chronic systemic damage due to the haematological phenotype.     

Circulating noradrenaline and β-adrenergic receptors in children with congestive heart failure

This study was designed to investigate changes in plasma catecholamine concentrations and the number of β-adrenoceptors (β-AR) of circulating lymphocyte in 94 noncyanotic congenital heart patients, in 43 patients with congestive heart failure, β-AR density was significantly lower ( p < 0.001) and plasma noradrenergic levels were significantly higher ( p < 0.001) compared with corresponding values in 51 patients without heart failure. A significant negative correlation between lymphocyte β-AR density and plasma noradrenergic levels was observed ( r =−0.61, p < 0.001). The degree of left-to-right shunt and pulmonary pressure was correlated directly with noradrenaline level and inversely with lymphocyte β-AR density. Both plasma noradrenaline level and lymphocyte β-AR density return to normal in children with heart failure after surgical repair. Our results support the idea that changes in noradrenaline level and lymphocyte β-adrenoceptor density occur concurrently with the presence and severity of heart failure in children.     

Serum interleukin-1β in neonatal sepsis

Serum levels of interleukin-1β (IL-1β) in newborn infants with septicaemia were measured and possible relationships between the clinical course of the infants, causative micro-organisms and IL-1β levels were investigated in a prospective study. The study groups comprised 49 newborn infants (25 mature, 24 premature) with proven sepsis and 40 healthy newborn infants (20 mature, 20 premature). Serum IL-1β levels were measured using the IL-1β immunoradiometric assay. The levels were found to be lower in neonates with sepsis (median 0.1 pg/ml) than in healthy controls (median 27.9 pg/ml) ( p <0.001). Non-significant trends towards lower levels were observed in children with shock and in non-survivors. No correlation was found between IL-1β and postnatal age, gestational age or the study weight of the patients. There was no significant difference in the serum IL-1β level in septic patients infected with Gram-positive bacteria and those infected with Gram-negative bacteria. The results show that the concentration of IL-1β is significantly decreased in preterm and term neonates with sepsis.     

Interleukin-10 and transforming growth factor-β1 in cord blood: relationship with paternal allergy and cesarean section

Aim: To measure Interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) in cord blood and assess their relationship with parental allergy and perinatal characteristics.
Methods: In a neonatal care unit 212 consecutive full-term and appropriate for gestational age newborns were recruited. IL-10 and TGF-β1 levels were determined in cord blood by high sensitivity ELISA. Perinatal characteristics, mode of delivery and presence of allergy in parents were recorded.
Results: Out of 212 newborns, 136 were of non-allergic parents and 76 (35.8%) of one or both allergic parents. In newborns of allergic fathers median IL-10 levels tended to be lower (0.67 vs. 1.06 pg/mL, p = 0.07) and TGF-β1 levels were significantly lower (40.9 vs. 45.3 ng/mL, p = 0.008) than in newborns of non-allergic parents. Multiple general regression analysis showed that presence of paternal allergy (β=−0.19, p = 0.003) to be born by cesarean section (β=−0.21, p = 0.03) and younger gestational age (β= 0.14, p = 0.04) independently contributed to decrease TGF-β1 levels (multiple R = 0.38, p < 0.0001).
Conclusion: Paternal allergy and cesarean section are associated to decreased TGF-β1, which might be the mediator of the increased risk of atopy development. Cord blood IL-10 and TGF-β1 levels of our newborn series could be used as reference values for further studies on these relationships.