Differentiation of murine pre-B cell line by an adjuvant muramyl peptide via NF-kappa B activation

Muramyl dipeptide (MDP) induces NF-kappa B activation in the murine pre-B cell line 70Z/3, increases the expression of surface immunoglobulins, and potentiates the response to other inducers such as LPS or IL-1. In the present study we investigated whether NF-kappa B activation was related to the MDP-stimulated immunoglobulin expression. In a gel shift assay our results confirmed that MDP but not MDP(D,D), an adjuvant-inactive stereoisomer, could induce a kappa B-binding activity in 70Z/3 cells. The LPS or IL-1 induced NF-kappa B binding activity was increased in the presence of MDP but not of MDP(D,D). A mutant of the cell line called 1.3E2, defective in NF-kappa B activations by LPS, did not respond to MDP. The enhanced surface immunoglobulin expression induced in the wild type 70Z/3 cells by MDP alone or combined to LPS, IL-1 or IFN gamma was not obtained in this variant. The ability of various treatments to activate the kappa gene enhancer was quantitatively evaluated in cells transfected with a kappa-enhancer-luciferase expression plasmid. Treatment of transfected 70Z/3 cells with MDP resulted in a dose-dependent enhancement of luciferase activity, an additive effect to that induced by LPS or IL-1. Treatment of the defective variant transfected with the same construct did not result in luciferase expression after stimulation with the various agents. The transient transfection assays were used to compare the effectiveness of some MDP analogs. Two adjuvant-active compounds unable to enhance kappa light chain expression did not increase the basal response in the transfected 70Z/3 cells, indicating that NF-kappa B activation was not related to the adjuvant potency of MDP but correlated with the kappa induction.