Diversity of the anti-T-cell receptor immune response and its implications for T-cell vaccination therapy of multiple sclerosis.
More precise understanding of the immune response against T-cell receptors (TCRs) is a prerequisite for successful TCR vaccination therapy of multiple sclerosis and other neurological autoimmune diseases. We conducted a detailed analysis of a paradigmatic anti-TCR response, using synthetic TCR peptides and highly purified recombinant TCR V alpha and Vbeta variable chains for the selection of CD4+ T-cell lines from a healthy volunteer. The target TCR (designated TCR(HWBP-3)) was obtained from HWBP-3, an autologous CD4+ T-cell line specific for myelin basic protein. The V alpha and Vbeta chains of TCR(HWBP-3) were expressed in Escherichia coli and purified by Ni-chelate chromatography and SDS (sodium dodecyl sulphate) gel electrophoresis. Further, we synthesized a set of 13- to 22-mer peptides spanning the complementarity-determining regions (CDR) 1, 2 and 3 and the framework regions (FR) of the alpha and beta chains of TCR(HWBP-3). The TCR peptides and proteins were then used to select a panel of TCR-specific CD4+ T-cell lines from donor HW. Several T-cell lines cross-reacted with a recombinant V chain and synthetic peptide. Cross-reactive immunogenic TCR epitopes were identified in the FR1 and CDR3 regions of the TCR(HWBP-3) alpha chain and in the FR1, CDR1 and CDR2 regions of the TCR(HWBP-3) beta chain. The TCR proteins and peptides were recognized in the context of at least three different HLA-DR molecules [DR2a (DRB5*0101), DR2b (DRB1*1501) and DRB1*1401/DRB3*0202]. Notably, the majority of the TCR peptide-selected T-cell lines did not react with the full-length recombinant V chains, suggesting they recognize 'cryptic' determinants. Based on the diversity of the anti-TCR immune response, we suggest that candidate TCR peptides should be screened in vitro in functional experiments before they are clinically applied for TCR vaccination therapy.
Department of Neuroimmunology, Max Planck Institute, Martinsried, Germany.
F
F
Zipp
M
M
Kerschensteiner
K
K
Dornmair
J
J
Malotka
S
S
Schmidt
A
A
Bender
G
G
Giegerich
R
R
de Waal Malefyt
H
H
Wekerle
R
R
Hohlfeld
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D000917:Antibody Formation / Q000502:physiology; D016207:Cytokines / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007167:Immunotherapy; D008285:Major Histocompatibility Complex / Q000276:immunology; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000628:therapy*; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism; D010641:Phenotype; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology* / Q000627:therapeutic use*; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology; D011994:Recombinant Proteins; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
D000906:Antibodies; D016207:Cytokines; D000939:Epitopes; D010446:Peptide Fragments; D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins