Immunoglobulin as a vehicle for foreign antigenic peptides immunogenic to T cells.
Antibody (Ab) molecules may serve as targeting vehicles for delivery of foreign antigenic peptides to antigen presenting cells (APC). An attractive strategy is to substitute segments between beta-strands of immunoglobulin (Ig) constant (C)-region domains with antigenic peptides. For this to work, the mutant Ab must maintain its conformation so that it can be secreted from transfected cells. Furthermore, the antigenic peptides must be excised by the processing machinery of APC and loaded onto major histo-compatibility complex (MHC) class II molecules. To test this, we have introduced a peptide of eleven amino acids (a.a.) as either of three different loops in the first C-region domain of the heavy (H) chain (CH1) of human IgG3. When the resulting mutant H chain genes were expressed in a fibroblast cell line equipped with proper class II molecules, the H chains were retained intracellularly, probably due to the light (L) chain deficiency of the fibroblasts. Nevertheless, by the endogenous class II processing pathway, presentation of the epitope to CD4+ cells was observed for all three mutants. The presentation efficiency, however, depended on the position of the peptide in the H chain. This could be due to influence of flanking sequences, which differ in the three loop replacement mutants. When L chain-expressing Chinese hamster ovary (CHO) lambda cells were transfected with the same constructs, two out of the three mutant Ig were secreted. The mutants had the expected antigen specificity and were recognized by anti-IgG Ab. When added exogenously to dendritic cell APC, the mutant IgG3 were processed, and the liberated foreign epitopes presented to T cells. The results suggest that the loops connecting beta-strands in the Ig fold may be replaced by foreign peptides, which upon processing become stimulatory to CD4+ T cells. Combined with the well-known targeting function of antibodies, this principle may be useful for construction of a new generation of vaccines.
Lunde
E
E
Institute for Immunology and Rheumatology, National Hospital, Oslo, Norway.
Bogen
B
B
Sandlie
I
I
D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D000941:Antigens / Q000276:immunology*; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D016466:CHO Cells; D006224:Cricetinae; D005803:Genes, Immunoglobulin; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D009154:Mutation; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D014162:Transfection
D000941:Antigens; D007136:Immunoglobulins; D011948:Receptors, Antigen, T-Cell