ACCUMULATION OF SOMATIC MUTANTS IN THE B-CELL COMPARTMENT AFTER PRIMARY IMMUNIZATION WITH A T-CELL-DEPENDENT ANTIGEN

The accumulation of somatic mutants in splenic B lymphocytes early after primary immunization with the hapten (4-hydroxy-3-nitro-phenyl)acetyl (NP) coupled to chicken gamma globulin (CG) was determined. Rearranged V186.2 heavy chain genes were amplified by the polymerase chain reaction from genomic DNA and subjected to nucleotide sequence analysis. Somatic antibody mutants become detectable on day 6 after immunization, and most of the somatic mutations accumulating in the memory compartment are introduced until day 14. At this time strong selection for mutants expressing high binding affinity for NP is apparent. Extrapolation from the mutation frequency increases between day 6 and day 14 to the previously determined mutation frequency at week 6 (Weiss, U. and Rajewsky, K., J. Exp. Med. 1990. 172: 1681) leads to the prediction that the process of mutant generation ceases to operate around day 22 after primary immunization.