Selection of a Single Antibody-Forming Cell Clone and Its Propagation in Syngeneic Mice

A single antibody-forming cell clone has been selected from primed mice by sequential transfer of limited numbers of spleen cells into irradiated syngeneic mice. The original spleen cell donors had been immunized with dinitrophenylated bovine gamma globulin. Specific antibody molecules in sera of recipient mice were separated by isoelectric focusing on polyacrylamide gels and visualized by 131 I-hapten binding and autoradiography. This method provided a marker for antibody-forming cells derived from a single cell clone. This report describes the history of one clone of cells (E9) producing IgG antibody to dinitrophenyl. Clone E9 is long-lived and has been maintained for five transplant generations (over 6 months) by serial transfer of spleen cells into irradiated syngeneic mice. Clone E9 has the following properties: (1) Antibody production strictly depends on antigen, presented either in vivo or in vitro; (2) Induction of E9 anti-dinitrophenyl shows specificity for the carrier protein; (3) Antibody is produced in amounts (2-3 mg/ml serum) comparable with myeloma-protein production by murine plasmacytomas; (4) In the absence of antigen, memory cells have a lifetime exceeding 28 days.