Targeting of influenza epitopes to murine CR1/CR2 using single-chain antibodies

• Published in: Immunopharmacology 1999-05, Vol.42 (1-3), p.159-165
• Main Authors: Prechl, József, Tchorbanov, Andrey, Horváth, Attila, Baiu, Dana C, Hazenbos, Wouter, Rajnavölgyi, Éva, Kurucz, István, Capel, Peter J.A, Erdei, Anna
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antigen targeting; Biological and medical sciences; Complement; Complement receptor 1; Complement receptor 2; complement receptors; Epitopes, B-Lymphocyte - genetics; Epitopes, B-Lymphocyte - immunology; Epitopes, T-Lymphocyte - genetics; Epitopes, T-Lymphocyte - immunology; Fundamental and applied biological sciences. Psychology; Fundamental immunology; Hemagglutinin Glycoproteins, Influenza Virus - immunology; Immunoglobulin Fragments - biosynthesis; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; Immunomodulators; Influenza A virus - immunology; influenza virus; Medical sciences; Mice; Molecular immunology; Molecular Sequence Data; Murine complement receptors type 1 and 2 (CR1/CR2); Pharmacology. Drug treatments; Rats; Receptors, Complement 3b - genetics; Receptors, Complement 3b - immunology; Receptors, Complement 3d - genetics; Receptors, Complement 3d - immunology; Recombinant Proteins - genetics; Recombinant Proteins - immunology; Single-chain variable fragment antibodies (scFv); Single-chain variable fragment antibody
• ISSN: 0162-3109
• DOI: 10.1016/S0162-3109(99)00025-9

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 IP1–37G6scFv 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 IP17G6 constructs as compared to the free peptide. The results suggest that scFv could work as targeting devices in subunit vaccines.