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. Author manuscript; available in PMC: 2015 Feb 5.

Published in final edited form as: J Control Release. 2010 Jul 7;148(2):219–225. doi: 10.1016/j.jconrel.2010.06.017

Fig. 3 — Enhanced functional cytolytic CD8⁺ T cells were detected in LLO-LPDII-primed mice by an in vivo CTL assay. (A) C57BL/6 mice were primed subcutaneously with PBS, 50 μg of OVA protein encapsulated in LLO-containing liposomes, 50 μg of pNGVL3.OVA in LLO-LPDII or in HI-LPDII, respectively. 12 days post-prime, all of the mice were boosted with 50 μg of OVA protein encapsulated in LLO-containing liposomes. 12 days post-boost, mice were i.v. injected with an equivalent amount of SIINFEKL-pulsed (labeled with 4 μM CFSE; CFSE^high) and non-pulsed (labeled with 0.4 μM CFSE; CFSE^low) splenocytes obtained from syngeneic naive donor mice. 16 hr after adoptive transfer of CFSE^high and CFSE^low cells, spleen cells from the recipient mice were harvested, and the proportions of the CFSE^high and CFSE^low cells were analyzed by flow cytometry. Representative flow cytometry data show counts of remaining non-pulsed CFSE^low splenocytes (left peak) versus remaining SIINFEKL-pulsed CFSE^high splenocytes (right peak) in histograms for naive, PBS prime/ protein boost, LLO-LPDII DNA prime/ protein boost, and HI-LLO-LPDII DNA prime/ protein boost mice. (B) Percentage of antigen peptide (SIINFEKL)-specific cell lysis is shown. The mean of the percentage from each group was compared to that of the PBS-primed group and was statistically analyzed (n = 4; * p < 0.05)

Fig. 3 — Enhanced functional cytolytic CD8⁺ T cells were detected in LLO-LPDII-primed mice by an in vivo CTL assay. (A) C57BL/6 mice were primed subcutaneously with PBS, 50 μg of OVA protein encapsulated in LLO-containing liposomes, 50 μg of pNGVL3.OVA in LLO-LPDII or in HI-LPDII, respectively. 12 days post-prime, all of the mice were boosted with 50 μg of OVA protein encapsulated in LLO-containing liposomes. 12 days post-boost, mice were i.v. injected with an equivalent amount of SIINFEKL-pulsed (labeled with 4 μM CFSE; CFSE^high) and non-pulsed (labeled with 0.4 μM CFSE; CFSE^low) splenocytes obtained from syngeneic naive donor mice. 16 hr after adoptive transfer of CFSE^high and CFSE^low cells, spleen cells from the recipient mice were harvested, and the proportions of the CFSE^high and CFSE^low cells were analyzed by flow cytometry. Representative flow cytometry data show counts of remaining non-pulsed CFSE^low splenocytes (left peak) versus remaining SIINFEKL-pulsed CFSE^high splenocytes (right peak) in histograms for naive, PBS prime/ protein boost, LLO-LPDII DNA prime/ protein boost, and HI-LLO-LPDII DNA prime/ protein boost mice. (B) Percentage of antigen peptide (SIINFEKL)-specific cell lysis is shown. The mean of the percentage from each group was compared to that of the PBS-primed group and was statistically analyzed (n = 4; * p < 0.05)