One-step cloning of anti tissue transglutaminase scFv from subjects with celiac disease.
Celiac disease is characterized by intestinal mucosal injury and malabsorption precipitated by dietary exposure to gluten of some cereals with a prominent role being played by gliadins, specific antigenic determinants found in wheat gluten. Patients suffering from celiac disease have serum antibodies recognizing gliadin, as well as the endomysial autoantigen tissue transglutaminase. Phage display antibody libraries have revealed ectopic production of anti-transglutaminase antibodies by intestinal lymphocytes with a biased use of the VH5 antibody gene family. Here we report a study on the pairing of VH and VL families in the antibodies to transglutaminase. Our results led to the construction of small phage display antibody libraries based on the amplification of the two genes in the VH5 family from intestinal lymphocytes. This method can be used for the rapid characterization of the anti-transglutaminase response in a potentially large number of subjects including asymptomatic patients whose serum antibodies may be undetectable.
Sblattero
Daniele
D
Department of Biology, University of Trieste, Via L. Giorgieri 10, 34127 Trieste, Italy. daniele@icgeb.org
Florian
Fiorella
F
Azzoni
Elisabetta
E
Ziberna
Fabiana
F
Tommasini
Alberto
A
Not
Tarcisio
T
Ventura
Alessandro
A
Bradbury
Andrew
A
Marzari
Roberto
R
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D001323:Autoantibodies / Q000235:genetics*; D002446:Celiac Disease / Q000201:enzymology / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular / Q000379:methods*; D019204:GTP-Binding Proteins / Q000276:immunology*; D005784:Gene Amplification; D006801:Humans; D007070:Immunoglobulin A / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007413:Intestinal Mucosa / Q000737:chemistry; D008214:Lymphocytes / Q000737:chemistry / Q000276:immunology; D008969:Molecular Sequence Data; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases / Q000276:immunology*
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D007070:Immunoglobulin A; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D000091345:Protein Glutamine gamma Glutamyltransferase 2; D011503:Transglutaminases; D019204:GTP-Binding Proteins