Immunoproteomics of processed beef proteins reveal novel galactose‐α‐1,3‐galactose‐containing allergens

Apostolović, Danijela; Tran, Thi Anh Thu; Hamsten, Carl; Starkhammar, Maria; Veličković, Tanja Ćirković; Hage, Marianne van

doi:10.1111/all.12462

Cited by 78 publications

(72 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We used our study cohort to identify α‐gal‐carrying proteins in beef because to the best of our knowledge, only five sera from patients with α‐gal‐mediated delayed meat allergy have been employed to address this question so far 30. Those patients showed multiple but overall faint bands in immunoblots.…”

Section: Discussionmentioning

confidence: 99%

The quantity and quality of α-gal-specific antibodies differ in individuals with and without delayed red meat allergy

Kollmann

Nagl

Ebner³

et al. 2016

Allergy

View full text Add to dashboard Cite

BackgroundIgG to galactose‐α‐1,3‐galactose (α‐gal) are highly abundant natural antibodies (Ab) in humans. α‐Gal‐specific IgE Ab cause a special form of meat allergy characterized by severe systemic reactions 3–7 h after consumption of red meat. We investigated 20 patients who experienced such reactions and characterized their α‐gal‐specific IgE and IgG responses in more detail.Methodsα‐Gal‐specific IgE was determined by ImmunoCAP. IgE reactivity to meat extract and bovine gamma globulin (BGG) was assessed by immunoblotting and ELISA, respectively. In some experiments, sera were pre‐incubated with α‐gal or protein G to deplete IgG Ab. α‐Gal‐specific IgG1–4 Ab in individuals with and without meat allergy were assessed by ELISA.ResultsIn immunoblots, BGG was the most frequently recognized meat protein. Binding of IgE and IgG to BGG was confirmed by ELISA and completely abolished after pre‐incubation with α‐gal. Neither the depletion of autologous α‐gal‐specific IgG Ab nor the addition of α‐gal‐specific IgG Ab from nonallergic individuals changed the IgE recognition of BGG of meat‐allergic patients. Meat‐allergic patients showed significantly higher α‐gal‐specific IgG1 and IgG3 Ab than nonallergic individuals, whereas the latter showed significantly higher levels of α‐gal‐specific IgG4 Ab.ConclusionPatients with delayed meat allergy display IgE and IgG Ab that selectively recognize the α‐gal epitope on BGG. Their enhanced α‐gal‐specific IgE levels are accompanied by high levels of α‐gal‐specific IgG1 devoid of IgE‐blocking activity. This subclass distribution is atypical for food allergies and distinct from natural α‐gal IgG responses in nonallergic individuals.

show abstract

Section: Discussionmentioning

confidence: 99%

The quantity and quality of α-gal-specific antibodies differ in individuals with and without delayed red meat allergy

Kollmann

Nagl

Ebner³

et al. 2016

Allergy

View full text Add to dashboard Cite

show abstract

“…Bands in mass range of 5–10, 10–15 and 15–20 kDa were subjected to in gel digestion and MS analysis on EASY nanoLC II system coupled with LTQ Orbitrap XL (Thermo Fisher Scientific Inc., Bremen, Germany), previously calibrated with the ProteoMass ™ LTQ/FT-Hybrid ESI Positive Mode Cal Mix (MSCAL5 SUPELCO, Sigma-Aldrich) as described previously 19 . The experiments were performed in duplicate.…”

Section: Methodsmentioning

confidence: 99%

“…Some of these proteins remain stable to thermal processing, showing that the allergenic properties are preserved in cooked meat 19, 20 . The long delay of symptoms in α-Gal allergy raises the question whether α-Gal glycoproteins have a retained IgE reactivity and allergenic properties when they appear in the circulation after digestion.…”

Section: Introductionmentioning

confidence: 99%

Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides

Apostolović

Krstic

Mihailović

et al. 2017

Sci Rep

Self Cite

View full text Add to dashboard Cite

The mammalian carbohydrate galactose-α1,3-galactose (α-Gal) causes a novel form of food allergy, red meat allergy, where patients experience severe allergic reactions several hours after red meat consumption. Here we explored gastric digestion of α-Gal glycoproteins using an in vitro model. Bovine thyroglobulin (BTG), a typical α-Gal carrying glycoprotein, was digested with pepsin. The resulting peptides were characterised by SDS PAGE, immunoblot and ImmunoCAP using sera from 20 red meat allergic patients. During pepsinolysis of BTG, a wide range of peptide bands was observed of which 14 to 17 kDa peptides remained stable throughout the gastric phase. The presence of the α-Gal epitope on the obtained peptides was demonstrated by an anti-α-Gal antibody and IgE from red meat allergic patients. The α-Gal digests were able to inhibit up to 86% of IgE reactivity to BTG. Importantly, basophil activation test demonstrated that the allergenic activity of BTG was retained after digestion in all four tested patients. Mass spectrometry-based peptidomics revealed that these peptides represent mostly internal and C-terminal parts of the protein, where the most potent IgE-binding α-Gal residues were identified at Asn1756, Asn1850 and Asn2231. Thus allergenic α-Gal epitopes are stable to pepsinolysis, reinforcing their role as clinically relevant food allergens.

show abstract

“…More recently, evidence has come from a study examining beef extracts using 2D gel electrophoresis. The authors demonstrated seven alpha-gal containing IgE binding proteins, four of which survived heating the beef extract 17 . How alpha- gal is structurally expressed on red meat remains unclear.…”

Section: The Red Meat Connectionmentioning

confidence: 99%

The alpha-gal story: Lessons learned from connecting the dots

Steinke

Platts-Mills

Commins

2015

Journal of Allergy and Clinical Immunology

311

245

View full text Add to dashboard Cite

Anaphylaxis is a severe allergic reaction that can be rapidly progressing and fatal; thus, establishing the etiology of anaphylaxis is pivotal to long-term risk management. Our recent work has identified a novel IgE antibody response to a mammalian oligosaccharide epitope, galactose-alpha-1,3-galactose (alpha-gal). IgE to alpha-gal has been associated with two distinct forms of anaphylaxis: i) immediate onset anaphylaxis during first exposure to intravenous cetuximab, and ii) delayed onset anaphylaxis 3–6 hours after ingestion of mammalian food products (e.g., beef and pork). Results of our studies and those of others strongly suggest that tick bites are a cause, if not the only significant cause, of IgE antibody responses to alpha-gal in the southern, eastern and central United States, Europe, Australia and parts of Asia. Typical immune responses to carbohydrates are considered to be T cell-independent, while IgE antibody production is thought to involve sequential class-switching that requires input from T cells. Therefore, establishing the mechanism of the specific IgE antibody response to alpha-gal will be an important aspect to address as this area of research continues.

show abstract

Immunoproteomics of processed beef proteins reveal novel galactose‐α‐1,3‐galactose‐containing allergens

Cited by 78 publications

References 32 publications

The quantity and quality of α-gal-specific antibodies differ in individuals with and without delayed red meat allergy

The quantity and quality of α-gal-specific antibodies differ in individuals with and without delayed red meat allergy

Peptidomics of an in vitro digested α-Gal carrying protein revealed IgE-reactive peptides

The alpha-gal story: Lessons learned from connecting the dots

Contact Info

Product

Resources

About