Characterization of a Functional Null Cell Line Derived from (NZB x NZW)F1 Mice

Abstract

We attempted to establish a null cell line from NZB x NZW(B/W)F1 mice in order to investigate a regulatory role of null cells during polyclonal B cell activation in autoimmune diseases. NB2.2, a representative subclone of resulting null cell lines, was maintained in long-term tissue culture with 10% mouse ConA supernatant (MCAS). Interestingly, the cell free supernatant of the NB2.2 cells (NB-CFS) showed marked synergistic effects on IgM secretion by B cells induced by IL-5. In addition, NB-CFS had the ability to augment the production of autoantibodies against bromelain-treated mouse red blood cells (BrMRBC) and single-stranded DNA (ssDNA) by B cells induced by IL-5. To determine whether NB2.2 cells induce polyclonal B cell activation and autoantibody generation in vivo, BALB/c mice were injected with NB2.2 cells. The results showed that the level of anti-ssDNA antibodies in sera of BALB/c mice injected with NB2.2 cells was significantly higher than that of control BALB/c mice injected with FDC-P2 cells. In addition, splenic B cells from mice injected with NB2.2 cells significantly proliferated in vitro in response to IL-4 and IL-5, and produced anti-ssDNA antibodies in the presence of IL-5. These results suggest that NB2.2, a null cell line established from B/WF1 mice, produces mediators capable of promoting polyclonal B cell activation and inducing autoantibody secretion, and that this kind of null cell may play an important role in the pathogenesis of autoimmune diseases.