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. 2013 Oct 14;8(10):e77346. doi: 10.1371/journal.pone.0077346

Figure 4. Purified B cells mediate cooperation between HEL105–120 and OVA323–339-specific CD4 T cells.

Figure 4

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 HEL105–120 alone or both HEL105–120 and OVA323–339. 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 HEL105–12-specific cytokine-producing cells in the spleens, and draining popliteal lymph nodes (DLN), were enumerated by ELISpot, in mice given HEL105–120-pulsed B cells (H), or HEL105–120 and OVA323–339-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.