In the present studies we investigated the early steps in B cell activation and determined that activation could be separated from entry into cell cycle. Purified B cells from BALB/c nu/nu mice, activated under controlled conditions by affinity-purified rabbit IgG anti-mu or F(ab')2 fragments prepared from the same antibodies, were sorted according to size by flow cytometry. Approximately 80% of the B cells were small and were shown to be in Go state by quantitative RNA analysis and by [3H]uridine and [3H]thymidine incorporation studies. The sorted Go B cells, when again incubated in serum-free medium with the appropriate anti-mu preparations, remained in Go. However, Go B cells in the presence of anti-mu have undergone significant change(s), in as much as they were now able to proliferate in response to soluble mediators. The ability to develop a proliferative response to B cell-activating factor(s), acquired independently from entry into cell cycle, characterizes another important step in early B cell activation and also indicates that Go B cells characterized on the basis of cell size, density, and/or RNA content may be in an activated state.

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