Bovine Holo-Beta-Lactoglobulin Cross-Protects Against Pollen Allergies in an Innate Manner in BALB/c Mice: Potential Model for the Farm Effect

Abstract

The lipocalin beta-lactoglobulin (BLG) is a major protein compound in cow’s milk, and we detected it in cattle stable dust. BLG may be a novel player in the farm protective effect against atopic sensitization and hayfever. In previous studies, we demonstrated that only the ligand-filled holo-form of BLG prevented sensitization to itself. Here, we investigated whether holo-BLG could, in an innate manner, also protect against allergic sensitization to unrelated birch pollen allergens using a murine model. BALB/c mice were nasally pretreated four times in biweekly intervals with holo-BLG containing quercetin–iron complexes as ligands, with empty apo-BLG, or were sham-treated. Subsequently, mice were intraperitoneally sensitized two times with apo-BLG or with the unrelated birch pollen allergen apo-Bet v 1, adjuvanted with aluminum hydroxide. After subsequent systemic challenge with BLG or Bet v 1, body temperature drop was monitored by anaphylaxis imaging. Specific antibodies in serum and cytokines of BLG- and Bet v 1-stimulated splenocytes were analyzed by ELISA. Additionally, human peripheral blood mononuclear cells of pollen allergic subjects were stimulated with apo- versus holo-BLG before assessment by FACS. Prophylactic treatment with the holo-BLG resulted in protection against allergic sensitization and clinical reactivity also to Bet v 1 in an unspecific manner. Pretreatment with holo-BLG resulted in significantly lower BLG-as well as Bet v 1-specific antibodies and impaired antigen-presentation with significantly lower numbers of CD11c+MHCII+ cells expressing CD86. Pretreatment with holo-BLG also reduced the release of Th2-associated cytokines from Splenocytes in BLG-sensitized mice. Similarly, in vitro stimulation of PBMCs from birch pollen allergic subjects with holo-BLG resulted in a relative decrease of CD3+CD4+ and CD4+CRTh2 cells, but not of CD4+CD25+CD127− Treg cells, compared to apo-BLG stimulation. In conclusion, prophylactic treatment with holo-BLG protected against allergy in an antigen-specific and -unspecific manner by decreasing antigen presentation, specific antibody production and abrogating a Th2-response. Holo-BLG therefore promotes immune resilience against pollen allergens in an innate manner and may thereby contribute to the farm protective effect against atopic sensitization.

Allergens and their associated small molecule ligands—their dual role in sensitization

“…It has been proposed that retinoic acid or iron‐quercetin would bind to T‐cell and B‐cell receptors and suppress immunogenicity and allergenicity of Bos d 5 80,81 . Based on a study in a mouse model, it was also suggested that ligand‐bound Bos d 5 was able to protect against allergic sensitization to unrelated birch pollen allergens 82 …”

“… 80 , 81 Based on a study in a mouse model, it was also suggested that ligand‐bound Bos d 5 was able to protect against allergic sensitization to unrelated birch pollen allergens. 82 …”

Long-term benefits of targeted micronutrition with the holoBLG lozenge in house dust mite allergic patients

“…The concept of targeted micronutrition to correct micronutritional deficiencies in immune cells of allergic patients, the main attribute of mode-of-action of loaded beta-lactoglobulin (holoBLG), has been unravelled only recently [3][4][5][6]. There are multiple studies showing that atopy is associated with iron deficiencies [7][8][9][10][11] and/or vitamin deficiencies [12][13][14][15].…”

Targeted micronutrition for allergic patients—possible applications of a food for special medical purposes

“…In addition to in vitro experiments and mouse models [11,18], the efficacy of holo-BLG in allergic patients has so far been investigated in two clinical pilot trials. In a first randomised, double-blind, placebo-controlled trial, a total of 20 female patients K Targeted micronutrition for allergic patients-possible applications of a food for special medical purposes 151 mini-review with birch pollen allergy were enrolled and received holo-BLG or placebo daily for 6 months (including the birch pollen season).…”