In vitro stimulation of immune spleen cells enhances the number of anti-lipid A-producing hybridomas.
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.
Erich
T
T
Laboratory for Microbiology, University of Utrecht, The Netherlands.
Dekker
B
B
De Beer
M
M
Torensma
R
R
Verhoef
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D002459:Cell Fusion; D002478:Cells, Cultured; D006825:Hybridomas / Q000276:immunology*; D007115:Immunization Schedule; D007074:Immunoglobulin G / Q000276:immunology; D066298:In Vitro Techniques; D008050:Lipid A / Q000008:administration & dosage / Q000276:immunology*; D051379:Mice; D012486:Salmonella typhimurium / Q000276:immunology; D013154:Spleen / Q000276:immunology
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008050:Lipid A