Mapping immune responses to mRBP-3 1-16 peptide with altered peptide ligands.
OBJECTIVE
Experimental autoimmune uveoretinitis (EAU) can be induced in C57BL/6 mice (I-A(b)) using human retinoid-binding protein-3 (hRBP-3, previously IRBP) residues 1-20. This study of a truncated murine peptide (mRBP-3 1-16) was conducted to determine its pathogenic potential and to characterize partially its interaction with specific T cells.


METHODS
After immunization with mRBP-3 1-16 or hRBP-3 1-20, EAU was assessed by immunohistochemistry. The immune response was assessed by tritiated thymidine incorporation and cytokine production analyzed by enzyme-linked immunosorbent assay (ELISA). T-cell receptor (TCR)- and major histocompatibility complex (MHC)-binding of mRBP-3 1-16 was studied by modeling and by using altered peptide ligands (APLs) and T-cell clones.


RESULTS
mRBP-3 1-16 induced EAU in C57BL/6 mice, with severity and kinetics comparable to that after immunization with hRBP-3 1-20. T cells taken from mice immunized with mRBP-3 1-16 had a Th1 phenotype and proliferated in response to reactivation with mRBP-3 1-16, hRBP-3 1-20, or mRBP-3 1-16 APLs. mRBP-3 1-16 APLs elicited at least five distinct patterns of reactivity when tested with the mRBP-3 1-16-reactive T-cell clones.


CONCLUSIONS
mRBP-3 1-16 immunizes and causes EAU in C57BL/6 mice. The studies using T-cell clones and APLs demonstrate that the immune response to mRBP-3 1-16 is drawn from a diverse population of antigen-specific T cells with a Th1 phenotype. Modeling and analysis of clones indicate that nonpathogenic T cells of an mRBP-3 1-16-reactive T-cell line recognize the peptide in a single register.
Guyver
Carly J
CJ
Department of Cellular and Molecular Medicine, School of Medical Sciences, University of Bristol, United Kingdom.
Copland
David A
DA
Calder
Claudia J
CJ
Sette
Alessandro
A
Sidney
John
J
Dick
Andrew D
AD
Nicholson
Lindsay B
LB
D000818:Animals; D001327:Autoimmune Diseases / Q000209:etiology* / Q000276:immunology / Q000473:pathology; D016207:Cytokines / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D005136:Eye Proteins / Q000276:immunology*; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D007114:Immunization; D008024:Ligands; D008198:Lymph Nodes / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D010446:Peptide Fragments / Q000276:immunology*; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D012173:Retinitis / Q000209:etiology* / Q000276:immunology / Q000473:pathology; D012177:Retinol-Binding Proteins / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology; D014605:Uveitis / Q000209:etiology* / Q000276:immunology / Q000473:pathology; D000099214:Retinol-Binding Proteins, Interstitial
D016207:Cytokines; D000939:Epitopes; D005136:Eye Proteins; D000949:Histocompatibility Antigens Class II; D008024:Ligands; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D012177:Retinol-Binding Proteins; D000099214:Retinol-Binding Proteins, Interstitial; C474482:I-A(b) antigen, mouse