Recombinant antibodies as carrier proteins for sub-unit vaccines: influence of mode of fusion on protein production and T-cell activation.
A major objective in development of vaccines is the design of sub-unit vaccines with the ability to induce strong T-cell responses. For this purpose, T-cell epitopes have been genetically inserted into various carrier proteins. Ig molecules may be especially useful as vehicles for delivery of CD4(+) T-cell epitopes to antigen presenting cells (APC). We have previously replaced loop structures between beta-strands in the C(H)1 domain of human IgG3 with a defined 11 amino acids long, MHC class II-restricted T-cell epitope. In this report we have added the same T-cell epitope into loops in the C(H)1 domain of mouse IgG2b. The following major points can be made: (1) Loops can accommodate an elongation of at least 11 amino acids without disruption of the overall Ig structure and secretion. (2) The recombinant Ig molecules are processed by spleen APC and the epitopes that are released are presented to T-cells. (3) Site of integration influences efficiency of processing and presentation. (4) Elongation of two neighbouring loops reduces Ig secretion. Taken together, our present results indicate that IgG C(H)1 domains may be engineered to carry T-cell epitopes in loop structures between beta-strands, but not all loops may be equally suitable for this purpose.
Eidem
J K
JK
University of Oslo, Department of Molecular Cell Biology, P.O. Box 1050, 0316, Oslo, Norway.
Rasmussen
I B
IB
Lunde
E
E
Gregers
T F
TF
Rees
A R
AR
Bogen
B
B
Sandlie
I
I
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002352:Carrier Proteins / Q000008:administration & dosage* / Q000235:genetics / Q000276:immunology; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D007074:Immunoglobulin G / Q000008:administration & dosage / Q000737:chemistry / Q000235:genetics; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000008:administration & dosage*
D000906:Antibodies; D002352:Carrier Proteins; D017931:DNA Primers; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic