In vitro generated human monoclonal trinitrophenyl-specific B cell lines. Evidence that human and murine anti-trinitrophenyl monoclonal antibodies cross-react with Escherichia coli beta-galactosidase.
Stable human antigen-specific monoclonal B cell lines were established without prior in vivo immunization. This was accomplished by expanding the anti-trinitrophenyl (TNP) B cells in vitro with the antigen TNP-Brucella abortus and then immortalizing them with Epstein-Barr virus. Five anti-TNP clones were selected by sequential limiting dilution. All five anti-TNP clones secreted IgM kappa antibodies. When tested against a panel of self and environmental antigens, all five anti-TNP clones exhibited cross-reactivity with an Escherichia coli-derived beta-galactosidase. To determine whether this was a more general phenomenon, a panel of murine monoclonals were tested and found to bind to beta-galactosidase. It is therefore possible that human and murine anti-TNP beta cell responses reflect reactivity against an environmental antigen, namely an epitope present on E. coli-derived beta-galactosidase. This approach of expanding human antigen-specific B cells by antigen stimulation in vitro, with a T-independent hapten-carrier conjugate before Epstein-Barr virus transformation, may prove useful in the development of human monoclonals for therapeutic purposes.
Golding
B
B
Division of Blood and Blood Products, Federal Drug Administration, Bethesda, MD 20892.
Inghirami
G
G
Peters
E
E
Hoffman
T
T
Balow
J E
JE
Tsokos
G C
GC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D002460:Cell Line; D002472:Cell Transformation, Viral; D003429:Cross Reactions; D004926:Escherichia coli / Q000201:enzymology / Q000276:immunology; D005696:Galactosidases / Q000276:immunology*; D004854:Herpesvirus 4, Human; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology; D007145:Immunoglobulin kappa-Chains / Q000276:immunology; D051379:Mice; D009578:Nitrobenzenes / Q000276:immunology*; D014301:Trinitrobenzenes / Q000276:immunology*; D001616:beta-Galactosidase / Q000276:immunology*
D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D007075:Immunoglobulin M; D007145:Immunoglobulin kappa-Chains; D009578:Nitrobenzenes; D014301:Trinitrobenzenes; D005696:Galactosidases; D001616:beta-Galactosidase