Mimicking the humoral immune response in vitro results in antigen-specific isotype switching supported by specific autologous T helper cells: generation of human HIV-1-neutralizing IgG monoclonal antibodies from naive donors

European Journal of Immunology
L T ChinC A Borrebaeck

Abstract

Molecular and cellular requirements for antigen-specific isotype switch of human B cells have been investigated by mimicking signaling occurring in germinal centers. Peripheral blood mononuclear cells from healthy seronegative blood donors were first primary immunized in vitro, using a synthetic immunogen containing both a T and B cell epitope, which generated specific IgM-secreting B cells. We used the apex of the V3 loop of gp120 as B cell epitope linked to a promiscuous T helper epitope from tetanus toxin. In parallel, CD4+ T helper cell clones specific for the T epitope of the immunogen were established. In a secondary in vitro stimulation period, we co-cultured the antigen-specific T and B cells on CD32-transfected fibroblasts, together with an anti-CD40 monoclonal antibody. This resulted in isotype switching and human antigen-specific, IgG-secreting B cells were detected. This response was strictly dependent upon the presence of autologous T helper cells and the immunogen. Antigen-specific human B cells derived from this primary and secondary in vitro immunization were subsequently subjected to electrofield-induced somatic cell hybridization and hybridomas secreting human anti-V3 IgG monoclonal antibodies were isolated. O...Continue Reading

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Citations

Apr 20, 2004·Toxicology Letters·Danuta J Herzyk, Elizabeth R Gore
Jan 15, 1997·Journal of Immunological Methods·A ZafiropoulosC A Borrebaeck
Oct 12, 2011·The Journal of Experimental Medicine·Hua-Xin LiaoBarton F Haynes
Nov 16, 2001·Biochemical and Biophysical Research Communications·K BradyP N Moynagh
Sep 6, 2000·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·A AkessonC A Borrebaeck

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