title: Human monoclonal antibodies against a recombinant HIV envelope antigen produced by primary in vitro immunization. Characterization and epitope mapping.
abstract: Peripheral blood lymphocytes from healthy, HIV sero-negative blood donors have been in vitro immunized using penv9, a recombinant fragment of the envelope of HIV-1. This primary in vitro immunization followed by Epstein-Barr virus (EBV) transformation and somatic cell fusion subsequently gave rise to several specific anti-penv9 monoclonal antibodies (MO28, MO30 and MO43) of mu isotype. The hybridomas have been kept in culture for over 6 months and the antibody productivity for MO30 was measured to 18 micrograms x (24 hr x 10(6) cells)-1. The fine specificity of the antibodies was mapped by a peptide inhibition enzyme immunoassay, using overlapping synthetic pentadeca peptides covering the whole penv9. These human monoclonal antibodies exhibited a similar epitope specificity directed against a non-sequential determinant, including the amino acids 632-646, 677-681 and 687-691. This specificity is very rarely found in immune sera from seropositive patients and presently not reported in human monoclonal antibodies derived from in vivo immunized individuals, indicating that different antibody specificities can be obtained by the in vitro immunization technology. These human monoclonal antibodies did not neutralize HIV. The results presented here demonstrate the feasability of generating human monoclonal antibodies against HIV by primary in vitro immunizations, thereby avoiding the use of lymphocytes derived from infected patients when human monoclonal antibodies for therapeutic purposes are to be produced.
authors: Department of Biotechnology, Lund University, Sweden.
authors: M
authors: M
authors: Ohlin
authors: P A
authors: PA
authors: Broliden
authors: L
authors: L
authors: Danielsson
authors: B
authors: B
authors: Wahren
authors: J
authors: J
authors: Rosen
authors: M
authors: M
authors: Jondal
authors: C A
authors: CA
authors: Borrebaeck
mesh_terms: D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D002459:Cell Fusion; D002461:Cell Line, Transformed; D002478:Cells, Cultured; D000939:Epitopes / Q000032:analysis*; D015686:Gene Products, env / Q000276:immunology*; D015488:HIV Antigens / Q000276:immunology*; D019302:HIV Envelope Protein gp160; D006801:Humans; D007114:Immunization / Q000379:methods; D008214:Lymphocytes / Q000276:immunology; D011498:Protein Precursors / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D015686:Gene Products, env; D015488:HIV Antigens; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors; D011994:Recombinant Proteins