Bitton N, Parizot C, Schindler D, Waks T, Debre P, Eshhar Z, Gorochov G; International Conference on AIDS.

Int Conf AIDS. 1996 Jul 7-12; 11: 62 (abstract no. Mo.A.1045).

Immunologie Cellulaire, CERVI, CNRS, Paris, France. Fax: (33 1) 42 17 74 90. E-mail: gorochov@ccr.jussieu.fr.

Objective: Engineering of T cells that express A): chimeric receptors with an antibody-type anti-HIV specificity that could eliminate infected cells in a non-MHc dependent fashion, or B): intracytoplasmic antibody fragments directed against the HIV-1 integrase. Methods: Human single chain Fvs (ScFvs) are selected from: random combinatorial naive antibody libraries or "disease" libraries constructed with genes originating from in vitro propagated and EBV transformed HIV patient's B cells. ScFvs are then expressed without leader sequence for intra-cellular immunisation or linked to the gamma chain of the FcgammaRIII for surface expression in T cell lines. Results: We have obtained several anti-ENV and anti-HIV-1 integrase antibodies under the ScFv format. Some of the anti-ENV ScFvs are able to bind the crude virus, and it is possible to select phage-displayed ScFvs using crude virus in order to obtain antibodies that bind oligomeric gp120. Chimeric receptors were then constructed under various formats with or without spacer regions (Ig Hinge, CD4 Ig domains, CD8 Hinge, Myc Tag) and compared in terms of optimal surface expression and antigen-induced cellular activation. In parallel, in vitro affinity maturation of the antibody fragment was obtained using either a chain shuffling approach or mutator strains. Conclusion: Phage display of antibody fragments appear to be particularly well suited for the selection and subsequent maturation of genes to be used in immunotherapy protocols.

Publication Types:

• Meeting Abstracts

Keywords:

• AIDS Vaccines
• Acquired Immunodeficiency Syndrome
• Animals
• Antibodies
• Antibody Specificity
• Antigens, CD4
• Bacteriophages
• Base Sequence
• HIV Envelope Protein gp120
• HIV Infections
• HIV Seropositivity
• Humans
• Immunoglobulins
• In Vitro
• Libraries
• T-Lymphocytes
• genetics
• immunology

Other ID:

• 96921116

UI: 102217015

From Meeting Abstracts