Ig isotype switching in B lymphocytes : A method for estimating isotype switch frequency in cloned B cell lymphomas
Whitmore, A. C.
Prowse, David M.
Arnold, L. W.
We have developed a method for enumerating the frequency of Ig isotype switching in clones of B cells. The method adapts Poisson statistics to analyze the distribution of amounts of switched isotype produced by multiple subclones of cells and thus enables one to estimate the probability that a single cell will switch isotype in one cell generation. We have applied this method to determine the spontaneous switch frequency of two Ly-1+ B cell lymphomas of B10-H-2aH-4bp/Wts mice. Both CH12.LX and CH27.LX switch from IgM to IgA at very high frequencies (1 - 5 x 10-3 switch events per cell division) and from IgM to IgG at low but detectable frequencies (10-4 - 10-5 switch events per cell division). Cloned IgG variants of CH12.LX switch to IgA at the same frequency as the IgM-producing cells. Bacterial lipopolysaccharide has a strong inhibitory effect on isotype switching by CH12.LX. Possible explanations for the observed preference for switching to IgA are discussed.