In vitro system for immunoglobulin class switching using BHK cells transfected with murine recombinant CD40 ligand

Komei Fukushima, Yoshiyuki Murata, Toshio Seto, Shuichi Furusawa, Haruo Matsuda

Research output: Contribution to journalArticlepeer-review


To develop an in vitro system for mouse immunoglobulin (Ig) class switching, the expression vector of murine CD40 ligand (CD40L) which is expressed on T cells was transfected to BHK cells. By using culture plates coated with the BHK cells expressing the recombinant CD40L, Ig class switching of splenic B cells was examined. The CD40L mRNA was cloned from splenic T cells of BALB/ mice activated with anti-CD3 antibody in vitro. As the No.593 base in the open reading frame sequence of the CD40L from BALB/c spleen differed from T to G, when compared with the known sequence from C57BL/6, one of the BALB/c-derived clones was reconstructed to the known CD40L by site-directed mutagenesis. Splenic B cells from BALB/c were induced secretion of Ig isotypes, IgM, IgG1 and IgE when cultured on two types of BHK cells, the transfected BHK cells with a CD40L clone from BALB/c and the transfected BHK cells with the reconstructed CD40L clone, in the presence of IL-4. However, when splenic B cells from C57BL/6 were cultured on the same systems, the B cells produced Ig isotypes, IgM, IgG1, IgG2a, IgG2b, IgG3 and IgE. In the similar experiments using the transfected BHK cells with a empty vector and the normal BHK cells, none of B cells produced any Ig isotypes other than IgM. These results indicate that Ig class switching of murine B cells can be induced by using these two types of CD40L-expressing BHK cells in vitro.

Original languageEnglish
Pages (from-to)615-619
Number of pages5
JournalJournal of Veterinary Medical Science
Issue number6
Publication statusPublished - Jun 1999
Externally publishedYes


  • BHK
  • CD40 ligand
  • Class switch
  • Immunoglobulin

ASJC Scopus subject areas

  • veterinary(all)


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