Abstract
An in vitro stimulation method for the generation of hybridomas producing antibodies with specificity for the weakly immunogenic lipid A is described. Conditions influencing in vitro stimulation of immune spleen cells were investigated. Depending on the experimental conditions the percentage of specific antibody-producing hybridomas varied between 0 and 39%. Most successful was stimulation with both antigen and the synthetic adjuvant muramyl dipeptide (MDP) for 3 days. In vitro stimulation of spleen cells from animals classically immunized with Salmonella Re mutant enhanced the number of lipid A-specific IgG-producing hybridomas from six after direct fusion to 17 after stimulation. These experiments indicate that the synergistic action of antigen and MDP is caused by preferential action on antigen selected B cells.
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