Purified B cells mediate cooperation between HEL_105–120 and OVA_323–339-specific CD4 T cells.

<p>BALB/c mice were injected with 10⁵ MACS purified CD4+ DO11.10 T cells one day prior to injection. Splenocytes from BALB/c mice were cultured in the presence of HEL_105–120 alone or both HEL_105–120 and OVA_323–339. After harvesting and washing, the splenic B cells were isolated by negative selection. B cells were injected in proportion to the standard injection of 10⁷, 3.5×10⁶ per mouse. On day 9 after the first injection the HEL_105–12-specific cytokine-producing cells in the spleens, and draining popliteal lymph nodes (DLN), were enumerated by ELISpot, in mice given HEL_105–120-pulsed B cells (H), or HEL_105–120 and OVA_323–339-pulsed B cells (H+O). Each data point represents the number of specific ELISpot-forming cells in the lymphoid organ of a single mouse. P-values indicate the probability that the means of the indicated samples are not different as assessed by unpaired, two-tailed T-tests. Data presented are pooled values from three independent experiments.