Isolation of variants of mouse myeloma X63 that express changed immunoglobulin class

Abstract

We have used fluorescence-activated cell sorting with class-specific antisera to isolate spontaneous variants in the expression of immunoglobulin heavy chain class from the mouse myeloma cell line X63 (IgGI, kappa). In the wild-type cell population, only one type of variants was found, namely, cells expressing IgG2b. From an IgG2b variant clone we isolated secondary variants that had either reverted to IgGI expression or expressed IgG2a or IgG2a and IgG2b concomitantly. The variant heavy chains are of normal size. The variant immunoglobulins were characterized serologically, and all of them still expressed the wild-type idiotype. Wild-type and variant cell populations were screened for heavy chain class-switch variants by fluorescence microscopy. A variety of switch variants was found in addition to the ones isolated by cell sorting, and a clear pattern of class switching (gamma 1 leads to gamma 2b leads to gamma 2a leads to alpha) with frequent reversion emerges from this analysis. Cells expressing new heavy chain classes occurred at frequencies of about 10(-7)-10(-6)/cell per generation, whereas revertants were as frequent as 10(-6)-10(-5)/cell per generation.