T cell recognition of carbohydrates on type II collagen.
A critical event in an immune response is the T cell recognition of peptides bound to major histocompatibility complex (MHC) molecules on the surface of an antigen presenting cell (APC). Although the majority of eukaryotic proteins are glycosylated, it has not yet been shown that T cell recognition of such proteins involves recognition of the bound carbohydrates. Type II collagen (CII), the major protein constituent of joint cartilage, is posttranslationally modified by hydroxylation and glycosylation of lysines. In this report we show that posttranslational modifications of the immunodominant peptide CII(256-270) generate a structural determinant that is distinct from the determinant represented by the corresponding synthetic peptide. Elimination of carbohydrates, present on CII, by two different biochemical methods revealed that the carbohydrates, O-linked to the hydroxylysines within the CII(256-270) determinant, were crucial for the reactivity towards the posttranslationally modified peptide. Furthermore, a T cell hybridoma specific for the glycosylated determinant was stimulated by tryptic CII-peptides presented by fixed APCs, thus showing that the carbohydrates are involved in the trimolecular complex T cell receptor/peptide/MHC. Finally, the importance of the bound carbohydrates for the arthritogenicity of CII was investigated by comparing the development of arthritis after immunization with carbohydrate-depleted and glycosylated CII, respectively. Incidence, time of onset, and severity of the disease were significantly affected by the elimination of carbohydrates, whereas no significant difference in anti-CII antibody titers was seen.
Michaëlsson
E
E
Department of Medical Inflammation Research, Lund University, Sweden.
Malmström
V
V
Reis
S
S
Engström
A
A
Burkhardt
H
H
Holmdahl
R
R
D000595:Amino Acid Sequence; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology; D002241:Carbohydrates / Q000276:immunology*; D003094:Collagen / Q000276:immunology* / Q000378:metabolism; D006825:Hybridomas / Q000276:immunology; D008285:Major Histocompatibility Complex / Q000276:immunology; D008969:Molecular Sequence Data; D011499:Protein Processing, Post-Translational; D051381:Rats; D013601:T-Lymphocytes / Q000276:immunology*
D002241:Carbohydrates; D003094:Collagen