Figure 4.

(a) The unresponsive clones do not proliferate to Hb(64–76) after one passage in culture. The anergized clones were “passed” once in culture (as described in Materials and Methods) and proliferation was again assessed (as described in Materials and Methods). (b) The anergic clones I + 10 μM 74L and I + 0.1 μM wt remain unresponsive to wt antigen after a period of 5 mo in culture. The clones were maintained in culture through a series of biweekly restimulations with antigen and APCs. 5 mo later, the clones were stimulated in vitro with Hb(64–76) and APCs. Proliferation was measured in a proliferation assay, as described.