首页 | 本学科首页   官方微博 | 高级检索  
检索        


Induction of antigen-specific isotype switching by in vitro immunization of human naive B lymphocytes
Authors:Alexandros Zafiropoulos  Eva Andersson  Elias Krambovitis  Carl A K Borrebaeck  
Institution:

a Department of Immunotechnology, Lund University, Lund, Sweden

b Laboratory of Applied Biochemistry and Immunology, Institute of Molecular Biology and Biotechnology, Heraklion, Greece

Abstract:The use of in vitro immunization technology for the generation of human antigen-specific antibodies has essentially resulted in low affinity IgM antibodies, resembling an in vivo primary immune response. We now describe a detailed reproducible protocol for a two-step in vitro immunization, which yields isotype switched, antigen-specific human antibodies. The immunizing antigen was a 30aa synthetic peptide, containing both a B (15aa V3 peptide of the HIV-1) and a T helper cell epitope (15aa peptide from tetanus toxin). The immunization protocol includes: (i) a selection procedure of donors with a memory T cell response against tetatus toxoid; (ii) immunization of mature naive peripheral B lymphocytes in two distinct phases, involving a primary and a secondary step. None of the donors which were examined after primary 7immunization showed at any time an IgG anti-V3 specific antibody response, while all the donors showed an IgM response. After the secondary immunization step, anti-V3 antibodies of both IgM and IgG isotypes were detected. The switch frequency event was high among the tested donors (5/8).
Keywords:In vitro immunization  Isotype switch  V3 loop peptide  Antibody  human  IgG  Tetanus toxoid  Helper epitope
本文献已被 ScienceDirect 等数据库收录!
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号