Analysis of cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulin

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Author(s)

Abstract

Although various in vitro immunization methods to generate antigen-specific antibodies have been described, a highly effective method that can generate high-affinity immunoglobulins has not yet been reported. Herein, we analyzed a cellular phenotype during in vitro immunization of murine splenocytes for generating antigen-specific immunoglobulins. We identified a combination of T cell-dependent stimuli (IL-4, IL-5, anti-CD38 and anti-CD4O antibodies) plus lipopolysaccharides (LPS) that stimulates antigen-exposed splenocytes in vitro, followed by induction of the cells phenotypically equivalent to germinal center B cells. We also observed that LPS induced high expression levels of mRNA for activation-induced cytidine deaminase. We stimulated antigen-exposed splenocytes, followed by the accumulation of mutations in immunoglobulin genes. From the immunized splenocytes, hybridoma clones secreting antigen-specific immunoglobulins were obtained.

Journal

  • Journal of bioscience and bioengineering

    Journal of bioscience and bioengineering 115(3), 339-345, 2013-03-25

    The Society for Biotechnology, Japan

References:  34

Codes

  • NII Article ID (NAID)
    110009596111
  • NII NACSIS-CAT ID (NCID)
    AA11307678
  • Text Lang
    ENG
  • Article Type
    ART
  • ISSN
    13891723
  • NDL Article ID
    024361695
  • NDL Call No.
    Z53-S65
  • Data Source
    CJP  NDL  NII-ELS  Crossref 
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