title: The industrial food additive, microbial transglutaminase, mimics tissue transglutaminase and is immunogenic in celiac disease patients.
abstract: Microbial transglutaminase (mTg) is capable of cross-linking numerous molecules. It is a family member of human tissue transglutaminase (tTg), and is involved in CD. Despite declarations of the safety of mTg for industrial use, direct evidence for immunogenicity of the enzyme is lacking. The serological activity of mTg, tTg, gliadin complexed mTg (mTg neo-epitope) and gliadin complexed tTg (tTg neo-epitope) were studied in 95 pediatric celiac patients (CD), 99 normal children (NC), 79 normal adults (NA) and 45 children with nonspecific abdominal pain (AP). Sera were tested by ELISAs, detecting IgA, IgG or both IgA and IgG (check): AESKULISA® tTg (tTg), AESKULISA® tTg New Generation (tTg neo-epitope (tTg-neo)), microbial transglutaminase (mTg) and mTg neo-epitope (mTg-neo). Marsh criteria were used for the degree of intestinal injury. Parallel, mTg and tTg neo-epitopes were purified by asymmetric field flow fractionation, confirmed by multi-light-scattering and SDS-PAGE, and analyzed in adult CD and control groups by competition ELISAs. No sequence homology but active site similarity were detected on alignment of the 2 Tgs. Comparing pediatric CD patients with the 2 normal groups: mTg-neo IgA, IgG and IgA+IgG antibody activities exceed the comparable mTg ones (p<0.0001). All mTg-neo and tTg-neo levels were higher (p<0.001). tTg IgA and IgG+IgA were higher than mTg IgA and IgA+IgG (p<0.0001). The levels of tTg-neo IgA/IgG were higher than tTg IgA/IgG (p<0.0001). The sequential antibody activities best reflecting the increased intestinal damage were tTg-neo check>tTg-neo IgA≥mTg-neo IgG>tTg-neo IgG>mTg-neo check>mTg-neo IgA. Taken together, tTg-neo check, tTg-neo IgA and mTg-neo IgG correlated best with intestinal pathology (r2=0.6454, r2=0.6165, r2=0.5633; p<0.0001, p<0.0001, p<0.0001, respectively). Purified mTg-neo IgG and IgA showed an increased immunoreactivity compared to single mTg and gliadin (p<0.001) but similar immunoreactivity to the tTg-neo IgG and IgA ELISA. Using competition ELISA, the mTg neo-epitopes and tTg neo-epitopes have identical outcomes in CD sera both showing a decrease in optical density of 55±6% (p<0.0002). mTg is immunogenic in children with CD and, by complexing to gliadin, its immunogenicity is enhanced. Anti-mTg-neo-epitope IgG antibodies correlate with intestinal damage to a comparable degree as anti-tTg-neo IgA. mTg and tTg display a comparable immunopotent epitope. mTg-neo IgG is a new marker for CD. Further studies are needed to explore the pathogenic potential of anti-mTg antibodies in CD.
authors.affiliation: AESKU.KIPP Institute, Wendelsheim, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Matthias
authors.affiliation: AESKU.KIPP Institute, Wendelsheim, Germany.
authors.forename: P
authors.initials: P
authors.lastname: Jeremias
authors.affiliation: AESKU.KIPP Institute, Wendelsheim, Germany.
authors.forename: S
authors.initials: S
authors.lastname: Neidhöfer
authors.affiliation: AESKU.KIPP Institute, Wendelsheim, Germany; B. Rappaport School of Medicine, Technion-Israel Institute of Technology, Haifa, Israel. Electronic address: aaronlerner1948@gmail.com.
authors.forename: A
authors.initials: A
authors.lastname: Lerner
mesh_terms: D002446:Celiac Disease / Q000276:immunology*; D005503:Food Additives / Q000737:chemistry*; D005511:Food Handling; D006801:Humans; D011503:Transglutaminases / Q000737:chemistry / Q000276:immunology*
chemical_list: D005503:Food Additives; D011503:Transglutaminases
keywords: Antibodies; Celiac disease; Food additive; Food processing; Immunogenic; Immunoreactivity; Microbial transglutaminase; Tissue transglutaminase