In Vitro Cell-Mediated Immunologic Assay for Cow's Milk Allergy

The production of a lymphokine, the leukocyte-migration-inhibition factor (LIF), by peripheral blood lymphocytes in response to an in vitro challenge with bovine β-lactoglobulin was assayed in infants and children suspected of having allergy to cow's milk protein. of the patients studied, 24 had cow's milk allergy, 24 were normal control subjects, 18 had recovered from milk allergy, 10 were newborns, and 10 were babies suffering from acute gastroenteritis. All patients with milk allergy demonstrated significant LIF production in response to β-lactoglobulin (23.5% ± 6.4%). In the normal control subjects, LIF was 3.1% ± 4.3% (P < .0005). Only two of the 24 control subjects and two of the ten newborns had high-normal values bordering on the positive. None of the ten babies with acute gastroenteritis gave a positive response. Most of the children who had recovered from milk allergy and were ingesting cow's milk had negative assays. This cell-mediated immune assay is shown to be a reliable test for the diagnosis of sensitivity to milk protein in infants and children, and for determining dietary treatment and when this treatment can be safely terminated. In most cases, its use should eliminate the need for the potentially dangerous and ethically questionable provocation test, as well as the need for repeated intestinal biopsies.

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Kinetics of in Vitro Response to β-Lactoglobulin in Children Allergic to and Tolerant of Cow’s Milk Protein

Journal of Investigational Allergology and Clinical Immunology ◽

10.18176/jiaci.0074 ◽

2016 ◽

Vol 26 (4) ◽

pp. 275-277

Author(s):

L Chini ◽

V Pacciani ◽

S Corrente ◽

E Monteferrario ◽

ML Romiti ◽

...

Keyword(s):

Milk Protein ◽

Cow’S Milk ◽

Cow's Milk ◽

In Vitro Response ◽

Cow’S Milk Protein ◽

Β Lactoglobulin ◽

Kinetics Of

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Allergen-responsive CD4+CD25+ Regulatory T Cells in Children who Have Outgrown Cow's Milk Allergy

Journal of Experimental Medicine ◽

10.1084/jem.20032121 ◽

2004 ◽

Vol 199 (12) ◽

pp. 1679-1688 ◽

Cited By ~ 343

Author(s):

Malin R. Karlsson ◽

Jarle Rugtveit ◽

Per Brandtzaeg

Keyword(s):

T Cells ◽

Treg Cells ◽

Cow’S Milk Allergy ◽

Cow’S Milk ◽

Milk Allergy ◽

Cow's Milk ◽

Cow's Milk Allergy ◽

Dietary Antigens ◽

Β Lactoglobulin

Cow's milk allergy in children is often of short duration, which makes this disorder an interesting clinical model for studies of tolerance to dietary antigens. Here, we studied T cell responses in 21 initially allergic children who, after a milk-free period of >2 mo, had cow's milk reintroduced to their diet. Children who outgrew their allergy (tolerant children) had higher frequencies of circulating CD4+CD25+ T cells and decreased in vitro proliferative responses to bovine β-lactoglobulin in peripheral blood mononuclear cells (PBMCs) compared with children who maintained clinically active allergy. No significant difference in proliferative activity stimulated by the polyclonal mitogen phytohemagglutinin was observed between the two groups. Depletion of CD25+ cells from PBMCs of tolerant children led to a fivefold increase in in vitro proliferation against β-lactoglobulin. This suggests that tolerance is associated with the appearance of circulating CD4+CD25+ regulatory T (Treg) cells that are capable of suppressing the effector T cells generated 1 wk after reintroduction of cow's milk. The suppressive function of the CD4+CD25+ Treg cells was shown to be partly cell contact dependent. Collectively, our study provides human data to suggest that mucosal induction of tolerance against dietary antigens is associated with the development of CD4+CD25+ Treg cells.

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Allergenicity of milk protein hydrolysate formulae in children with cow's milk allergy

European Journal of Pediatrics ◽

10.1007/bf01953996 ◽

1993 ◽

Vol 152 (9) ◽

pp. 760-762 ◽

Cited By ~ 103

Author(s):

V. Ragno ◽

P. G. Giampietro ◽

G. Bruno ◽

L. Businco

Keyword(s):

Milk Protein ◽

Protein Hydrolysate ◽

Cow’S Milk Allergy ◽

Cow’S Milk ◽

Milk Allergy ◽

Cow's Milk ◽

Cow's Milk Allergy

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Human Breast Milk Contains Bovine lgG. Relationship to Infant Colic?

PEDIATRICS ◽

10.1542/peds.87.4.439 ◽

1991 ◽

Vol 87 (4) ◽

pp. 439-444

Author(s):

Patrick S. Clyne ◽

Anthony Kulczycki

Keyword(s):

Breast Milk ◽

Human Milk ◽

Milk Protein ◽

Cow’S Milk ◽

Human Breast Milk ◽

Human Breast ◽

Cow's Milk ◽

Milk Samples ◽

Infant Colic ◽

Β Lactoglobulin

Previous studies have suggested that an unidentified cow's milk protein, other than β-lactoglobulin and casein, might play a pathogenetic role in infant colic. Therefore, a radioimmunoassay was used to analyze human breast milk and infant formula samples for the presence of bovine IgG. Milk samples from 88 of the 97 mothers tested contained greater than 0.1 µg/mL of bovine IgG. In a study group of 59 mothers with infants in the colic-prone 2- to 17-week age group, the 29 mothers of colicky infants had higher levels of bovine IgG in their breast milk (median 0.42 µg/mL) than the 30 mothers of noncolicky infants (median 0.32 µg/mL) (P < .02). The highest concentrations of bovine IgG observed in human milk were 8.5 and 8.2 µg/mL. Most cow's milk-based infant formulas contained 0.6 to 6.4 µg/mL of bovine IgG, a concentration comparable with levels found in many human milk samples. The results suggest that appreciable quantities of bovine IgG are commonly present in human milk, that significantly higher levels are present in milk from mothers of colicky infants, and that bovine IgG may possibly be involved in the pathogenesis of infant colic.

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Colic or Milk Allergy

PEDIATRICS ◽

10.1542/peds.84.5.938 ◽

1989 ◽

Vol 84 (5) ◽

pp. 938-939

Author(s):

BRUCE TAUBMAN

Keyword(s):

Milk Protein ◽

Cow’S Milk ◽

Milk Allergy ◽

Cow's Milk ◽

Healthy Infants ◽

Infant Colic ◽

Excessive Crying ◽

Cow’S Milk Protein ◽

Definition Of ◽

The Relationship

In the article by Drs Lothe and Lindberg, they purport to examine the relationship between infant colic and cow's milk protein allergy. However, the majority of the infants they studied did not fit the definition of infants with colic. The authors claimed the definition they used for their study corresponded to that of Wessels et al, Carey, and Schmitt. Yet, each of these physicians defined the syndrome of infant colic as excessive crying in "otherwise healthy" infants.

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Cow's Milk Allergy in Infants and Children

Journal of Nutritional Medicine ◽

10.3109/13590849109084116 ◽

1991 ◽

Vol 2 (2) ◽

pp. 201-201

Author(s):

Norman W. Clein

Keyword(s):

Infants And Children ◽

Cow’S Milk Allergy ◽

Cow’S Milk ◽

Milk Allergy ◽

Cow's Milk ◽

Cow's Milk Allergy ◽

In Infants And Children

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Peptide Release after Simulated Infant In Vitro Digestion of Dry Heated Cow’s Milk Protein and Transport of Potentially Immunoreactive Peptides across the Caco-2 Cell Monolayer

Nutrients ◽

10.3390/nu12082483 ◽

2020 ◽

Vol 12 (8) ◽

pp. 2483 ◽

Cited By ~ 1

Author(s):

Hannah E. Zenker ◽

Harry J. Wichers ◽

Monic M. M. Tomassen ◽

Sjef Boeren ◽

Nicolette W. De Jong ◽

...

Keyword(s):

T Cell ◽

Milk Protein ◽

Cell Monolayer ◽

In Vitro Digestion ◽

Cow’S Milk ◽

T Cell Epitopes ◽

Peptide Profile ◽

Cow's Milk ◽

Cow’S Milk Protein

Dry heating of cow’s milk protein, as applied in the production of “baked milk”, facilitates the resolution of cow’s milk allergy symptoms upon digestion. The heating and glycation-induced changes of the protein structure can affect both digestibility and immunoreactivity. The immunological consequences may be due to changes in the peptide profile of the digested dry heated milk protein. Therefore, cow’s milk protein powder was heated at low temperature (60 °C) and high temperature (130 °C) and applied to simulated infant in vitro digestion. Digestion-derived peptides after 10 min and 60 min in the intestinal phase were measured using LC-MS/MS. Moreover, digests after 10 min intestinal digestion were applied to a Caco-2 cell monolayer. T-cell epitopes were analysed using prediction software, while specific immunoglobin E (sIgE) binding epitopes were identified based on the existing literature. The largest number of sIgE binding epitopes was found in unheated samples, while T-cell epitopes were equally represented in all samples. Transport of glycated peptide indicated a preference for glucosyl lysine and lactosyl-lysine-modified peptides, while transport of peptides containing epitope structures was limited. This showed that the release of immunoreactive peptides can be affected by the applied heating conditions; however, availability of peptides containing epitopes might be limited.

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Effect of Low-Immunogenic Yogurt Drinks and Probiotic Bacteria on Immunoreactivity of Cow’s Milk Proteins and Tolerance Induction—In Vitro and In Vivo Studies

Nutrients ◽

10.3390/nu12113390 ◽

2020 ◽

Vol 12 (11) ◽

pp. 3390

Author(s):

Barbara Wróblewska ◽

Anna Kaliszewska-Suchodoła ◽

Ewa Fuc ◽

Lidia Hanna Markiewicz ◽

Anna Maria Ogrodowczyk ◽

...

Keyword(s):

Therapeutic Potential ◽

Tolerance Induction ◽

Milk Proteins ◽

In Vivo Studies ◽

Cow’S Milk ◽

Milk Allergy ◽

Cow's Milk ◽

Safe Product

There is no effective therapy for milk allergy. The role of lactic acid bacteria (LAB) and probiotics in protection against allergy-related outcomes is still under investigation. The aim of the study was to evaluate the immunomodulative and therapeutic potential of yogurt drinks in cow’s milk allergy (CMA) management. We compared immunoreactivity of α-casein (α-CN), β-casein (β-CN), κ-casein (κ-CN), α-lactalbumin (α-LA), and β-lactoglobulin (β-LG) in 27 yogurt drinks fermented with different basic yogurt cultures, or yogurt cultures enriched with Lactobacillus plantarum and/or Bifidobacterium lactis strains, by competitive ELISA assay. Drinks with the lowest antigenic potential were used as allergoids for CMA therapy. BALB/c mice were sensitized via intraperitoneal injection of α-CN + β-LG mixture with aluminum adjuvant, and gavaged with increasing doses of selected low-immunogenic drinks (YM—basic, or YM-LB—enriched with L. plantarum and B. lactis) to induce tolerance. Milk- or phosphate-buffered saline (PBS)-dosed mice served as controls. Compared to milk, the immunoreactivity of proteins in drinks increased or decreased, depending on the bacterial sets applied for fermentation. Only a few sets acted synergistically in reducing immunoreactivity. The selected low-immunogenic drinks stimulated allergic mice for profiling Th2 to Th1 response and acquire tolerance, and the effect was greater with YM-LB drink, which during long-lasting interventional feeding strongly increased the secretion of regulatory cytokines, i.e., IL-10 and TGF-β, and IgA and decreased IL-4, IgE, and anti-(α-CN + β-LG) IgG1. The studies revealed variations in the potency of yogurt bacteria to change allergenicity of milk proteins and the need for their strict selection to obtain a safe product for allergy sufferers. The YM-LB drink with reduced antigenic potential may be a source of allergoids used in the immunotherapy of IgE mediated CMA, but further clinical or volunteer studies are required.

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Individual Sensitization Pattern Recognition to Cow’s Milk and Human Milk Differs for Various Clinical Manifestations of Milk Allergy

Nutrients ◽

10.3390/nu11061331 ◽

2019 ◽

Vol 11 (6) ◽

pp. 1331 ◽

Cited By ~ 4

Author(s):

Frauke Schocker ◽

Skadi Kull ◽

Christian Schwager ◽

Jochen Behrends ◽

Uta Jappe

Keyword(s):

Human Milk ◽

Clinical Manifestations ◽

Basophil Activation Test ◽

Food Allergies ◽

Cow’S Milk ◽

Human Breast Milk ◽

Milk Allergy ◽

Cow's Milk ◽

Single Allergens

Cow’s milk allergy (CMA) belongs to one of the most common food allergies in early childhood affecting 2–3% of children under 3 years of age. However, approximately 1% of adults remain allergic to cow’s milk, often showing severe reactions even to traces of milk. In our study, we recruited patients with different clinical manifestations of CMA, including patients with anaphylaxis and less severe symptoms. We assessed the sensitization patterns and allergic responses of these subgroups through different immunological and cell-based methods. Sera of patients were investigated for IgE against whole cow’s milk and its single allergens by CAP- FEIA. In a newly developed in-house multiplex dot assay and a basophil activation test (BAT), cow’s milk allergens, in addition to human breast milk and single allergens from cow’s and human milk were analyzed for IgE recognition and severity of CMA in the included patients. Both the CAP-FEIA routine diagnostic and the multiplex dot test could differentiate CMA with severe from milder allergic reactions by means of the patients’ casein sensitization. The BAT, which mirrors the clinical response in vitro, confirmed that basophils from patients with severe reactions were more reactive to caseins in contrast to the basophils from more moderate CMA patients. By means of this improved component-resolved diagnosis of CMA, individual sensitization patterns could be assessed, also taking sensitization against human milk into consideration.

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