Monoclonal antibodies to murine CD40 define two distinct functional epitopes

Eur J Immunol. 1994 Aug;24(8):1828-34. doi: 10.1002/eji.1830240816.

Abstract

Two rat IgG2a antibodies which define distinct epitopes on murine CD40 have been generated. These antibodies specifically bind recombinant murine CD40 expressed on L cells, and the soluble extracellular domain of murine CD40 coated onto microtiter plates. Both antibodies bind B220+ but not B220 murine spleen cells, and immunoprecipitate a 45-kDa protein from the surface of purified murine splenic B cells. These antibodies exhibit separate functional properties, consistent with the notion that they define two distinct CD40 epitopes. One of the monoclonal antibodies (designated 1C10) directly induces a specific proliferative response from mature murine B cells, up-regulates several B cell surface antigens, and rescues immature B lymphoma cells from anti-IgM-induced growth arrest. The other monoclonal antibody (designated 4F11) exhibits none of these properties, but is capable of synergizing with suboptimal amounts of either anti-IgM antibodies or the 1C10 agonistic anti-CD40 antibody to produce an optimal proliferative response of purified small dense B cells. Furthermore, 4F11 antibody synergizes with suboptimal amounts of 1C10 antibody to rescue B lymphoma cells from anti-IgM-induced growth arrest. The 1C10 and 4F11 antibodies were unable to cross-block each other's binding to recombinant CD40 expressed in L cells, providing strong support for the notion that the antibodies recognize distinct epitopes on CD40. The potential implications of two functionally distinct CD40 epitopes are discussed.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antigenic Modulation / immunology
  • Antigens, CD / immunology*
  • Antigens, Differentiation, B-Lymphocyte / immunology*
  • B-Lymphocytes / immunology
  • CD40 Antigens
  • Cell Division / immunology
  • Cross Reactions
  • Epitopes / immunology*
  • Female
  • Fluorescent Antibody Technique
  • Mice
  • Mice, Inbred BALB C
  • Precipitin Tests
  • Rats
  • Rats, Inbred Lew
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Antigens, CD
  • Antigens, Differentiation, B-Lymphocyte
  • CD40 Antigens
  • Epitopes