Specific reactivity of fluorescein isothiocyanate-conjugated separated IgG and IgA from celiac disease sera on human tissues.
Study of the reaction of celiac disease-related antibodies with human substrates has been hampered by the immunological cross-reactivity between fluorescein isothiocyanate-conjugated anti-human immunoglobulins and immunoglobulins normally present in human tissues. In order to overcome this problem we extracted IgG from 2 celiac disease sera (positive for specific IgG and IgA antibodies) using a genetically engineered recombinant form of streptococcal protein G covalently immobilized on 4% agarose beads. The separated IgG and IgA was conjugated with fluorescein isothiocyanate and used in direct immunofluorescence on 0 blood group human duodenum, liver, and myocardium. Staining of the lamina propria beneath the villous epithelium, the endomysium of smooth muscle layers in the duodenum, and the liver sinusoidal and pericellular myocardial matrix was observed. The reactivity of celiac disease-related antibodies with various human tissues indicates that these antibodies are the result of a systemic immune response directed against all tissues containing matrix proteins.
Volta
U
U
Istituto di Clinica Medica Generale e Terapia Medica, Università di Bologna, Policlinico S. Orsola, Italy.
Ballardini
G
G
Molinaro
N
N
De Franceschi
L
L
Groff
P
P
Bianchi
F B
FB
D000818:Animals; D000918:Antibody Specificity; D002446:Celiac Disease / Q000276:immunology*; D004386:Duodenum / Q000276:immunology; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D006801:Humans; D007070:Immunoglobulin A / Q000302:isolation & purification / Q000378:metabolism*; D007074:Immunoglobulin G / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000276:immunology; D009206:Myocardium / Q000276:immunology; D009928:Organ Specificity
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D016650:Fluorescein-5-isothiocyanate