Targeting of influenza epitopes to murine CR1/CR2 using single-chain antibodies.
Single-chain variable fragment (scFv) antibodies are genetically engineered molecules comprising the variable regions responsible for specific binding. scFv that recognize certain surface molecules on professional antigen presenting cells could therefore be suitable for targeting Ag to these cells. We have produced an scFv that recognizes murine complement receptors 1 and 2 (CR1/CR2) and genetically fused it with different numbers of influenza hemagglutinin peptides which contain both B and T cell epitopes. The CR1/CR2 specific hybridoma 7G6 was used for RT-PCR to obtain the variable regions, which were then combined to create an scFv fragment. The influenza hemagglutinin intersubunit peptide HA317-41 (IP) was engineered to the N terminus of the scFv in one, two or three copies. The so obtained IP(1-3)7G6scFv still bound the complement receptors; the peptides in the construct were recognized by the peptide specific monoclonal IP2-11-1 on Western blots and ELISAs. The CR1/CR2 positive B lymphomas A20 and 2PK3 presented the peptide to an I-Ed restricted IP specific T cell hybridoma more efficiently when incubated with the IP(1)7G6 constructs as compared to the free peptide. The results suggest that scFv could work as targeting devices in subunit vaccines.
Department of Immunology, L. Eötvös University, Göd Jávorka S.u., Hungary. jprechl@mail.datanet.hu
J
J
Prechl
A
A
Tchorbanov
A
A
Horváth
D C
DC
Baiu
W
W
Hazenbos
E
E
Rajnavölgyi
I
I
Kurucz
P J
PJ
Capel
A
A
Erdei
D000595:Amino Acid Sequence; D000818:Animals; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D018984:Epitopes, T-Lymphocyte / Q000235:genetics / Q000276:immunology*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D009980:Influenza A virus / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D017463:Receptors, Complement 3b / Q000235:genetics / Q000276:immunology*; D017464:Receptors, Complement 3d / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D017463:Receptors, Complement 3b; D017464:Receptors, Complement 3d; D011994:Recombinant Proteins